This invention relates to dihydroquinazoline derivatives, or pharmaceutically acceptable salts thereof, which possess anti-cancer activity. The invention includes dihydroquinazoline derivatives, processes for their manufacture, pharmaceutical compositions containing them and their use in the manufacture of a medicament for use in the treatment or prevention of cancer in a warm-blooded animal such as man.
It is known that inhibitors of enzymes which use folic acid derivatives often possess anti-cancer activity. Examples of such compounds include antimetabolites such as aminopterin and methotrexate. A newer compound of this type which showed considerable promise in clinical trials is known as CB3717 and is described in UK-B-2065653. Despite its promising activity against human breast, ovarian and liver cancer, however, CB3717 shows symptoms of toxicity in humans, particularly in relation to the liver and kidney (Cancer Treatment Reports, 1986, 70, 1335). A newer compound which possesses inhibitory activity against thymidylate synthase and which is showing promise in clinical trials is known as raltitrexed or ZD1694 (N-(5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-2-thenoyl)-L-glutamic acid) and is described in EP-A-239362. The compound has been shown to possess promising activity against human breast, ovarian, colorectal and non-small cell lung cancer and against adenocarcinoma (8th NCI-EORTC Symposium, Amsterdam, March, 1994; Abstract Nos. 240 and 242 to 245).
Compounds of the ZD1694 and CB3717-type are believed to act as anti-tumour agents by inhibiting the enzyme thymidylate synthase, which catalyses the methylation of deoxyuridine monophosphate to produce thymidine monophosphate which is required for DNA synthesis. The anti-cancer activity of ZD1694 and CB3717 may be assessed in vitro by determining its inhibitory effect on that enzyme, and in cell cultures by its inhibitory effect on cancer cell lines such as the mouse leukaemia cell line L1210, the mouse lymphoma cell line L5178Y TKxe2x88x92/xe2x88x92 and the human breast cancer cell line MCF-7.
Other compounds of the ZD1694 and CB3717-type have been described and claimed in EP-A-239362, EP-A-284338, EP-A-339976, EP-A-373891, EP-A-459730, EP-A-509643 and EP-A-562734, and in WO 93/19051, WO 94/07869 and WO 94/11354. These compounds may have their anti-cancer activity assessed by their activity against, for example, thymidylate synthase and the L1210, L5178Y TKxe2x88x92/xe2x88x92 and MCF-7 cell lines.
Although inhibitors of thymidylate synthase are useful in many conditions, there is a need for anti-cancer compounds which operate by a different mechanism. Such compounds are required for treating patients with cancers refractory to standard chemotherapeutic agents. Further, it is highly desirable that anti-cancer compounds are water soluble, as increased water solubility generally enables a higher bioavailability.
4-[N-[7-bromo-2-methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide (CB 30865) is a known anti-cancer compound which acts via a non-folate dependent locus (Skelton et al, Cytometry 33 56-66 (1998)). However, the compound has very low water-solubility and is not suitable for in vivo administration. The compound is also still a good inhibitor of isolated thymidylate synthase and in some cell lines (resistant to the primary effects of the compound) this becomes a growth rate limiting target for CB 30865 at compound concentrations in the micromolecular range.
The present invention therefore seeks to provide new anti-cancer compounds which do not operate via inhibition of thymidylate synthase and which have an acceptable bioavailability.
Accordingly, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, 
wherein:
either R1 and R1xe2x80x2 together form an oxo group and R2 is hydrogen, C1-C4 alkyl, xe2x80x94(C1-C4 alkyl)xe2x80x94B, xe2x80x94(C1-C4 alkyl)xe2x80x94COB, xe2x80x94(C1-C4 alkyl)xe2x80x94COxe2x80x94(C1-C4 alkyl)xe2x80x94B, xe2x80x94(C1-C4 alkyl)xe2x80x94CO2xe2x80x94(C1-C4 alkyl)xe2x80x94B, xe2x80x94(C1-C4 alkyl)xe2x80x94CO2xe2x80x94(C2-C4 alkenyl)xe2x80x94B or xe2x80x94(C1-C4 alkyl)xe2x80x94CONHxe2x80x94(C1-C4 alkyl)xe2x80x94B wherein B is xe2x80x94CO2H, hydroxy, C1-C4 alkoxy, amino, (C1-C4 alkyl)amino, di(C1-C4 alkyl)amino or a 5- or 6-membered heterocyclic group, or R1xe2x80x2 and R2 together form a bond and R1 is xe2x80x94Sxe2x80x94(C1-C4 alkyl), xe2x80x94NHRxe2x80x2 or xe2x80x94NHCORxe2x80x2 wherein Rxe2x80x2 is aryl or C1-C4 alkyl;
R3 is xe2x80x94(CH2)pxe2x80x94A wherein p is from 1 to 4 and A is a 5- or 6-membered N-containing heterocyclic ring attached via the N atom or A is xe2x80x94NAxe2x80x2Axe2x80x3 wherein Axe2x80x2 and Axe2x80x3 are the same or different and are each a C1-C4 alkyl group;
either R4 is hydrogen, oxo or C1-C4 alkyl and R5 is hydrogen, C1-C4 alkyl or halogen, or R4 and R5, together with the carbon atoms to which they are attached, form a 5- or 6-membered carbocyclic ring;
X1 is xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94 or xe2x80x94NRxe2x80x3xe2x80x94 wherein Rxe2x80x3 is hydrogen, C1-C4 alkyl, C2-C4 alkenyl or C2-C4 alkynyl;
Y is a divalent aryl or heteroaryl group;
R6 is hydrogen, oxo or C1-C4 alkyl;
X2 is xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94 or xe2x80x94NRxe2x80x3xe2x80x94 wherein Rxe2x80x3 is as defined above;
m is from 1 to 4; and
R7 is pyridyl, pyrimidyl, imidazolyl, triazolyl, xe2x80x94(C1-C4 alkyl)-imidazolyl, or xe2x80x94(C1-C4 alkyl)-triazolyl.
As used herein, a C1-C4 alkyl group or moiety can be linear or branched but is preferably linear. Suitable such alkyl groups and moieties include methyl, ethyl, n-propyl, i-propyl, n-butyl and t-butyl. Methyl and ethyl are preferred and methyl is particularly preferred. A C1-C4 alkyl group or moiety can be substituted or unsubstituted at any position. Typically, it carries up to 3 substituents, e.g. one or two substituents. Suitable substituents include hydroxy, halogen, amino, (C1-C4 alkyl)amino and di-(C1-C4 alkyl)amino. Preferred substituents are dimethylamino, diethylamino and halogen.
In groups which contain more than one C1-C4 alkyl moiety, the alkyl moieties may be the same or different. Thus, the C1-C4 alkyl moieties present in a di-(C1-C4 alkyl)amino group may be the same or different.
A C2-C4 alkenyl group is typically an allyl group.
An aryl group or moiety is typically a C6-C10 aryl group or moiety. Suitable such aryl groups and moieties include phenyl and naphthyl. Phenyl is preferred. An aryl group or moiety may be substituted or unsubstituted at any position. Preferably, it is unsubstituted. However, it can carry, for example, 1, 2, 3 or 4 substituents. Suitable substituents include halogen, hydroxyl, said C1-C4 alkyl such as methyl or ethyl, C1-C4 alkoxy such as methoxy or ethoxy, amino, (C1-C4 alkyl)amino and di-(C1-C4 alkyl)amino. A divalent aryl moiety is preferably a 1,4-phenylene group.
p is typically 1 or 2.
A halogen is typically chlorine, fluorine, bromine or iodine. Chlorine and bromine are preferred.
A said 5- or 6-membered carbocyclic ring may be a substituted or unsubstituted, aromatic or non-aromatic 5- or 6-membered ring made up of carbon atoms. Preferably, it is a 5-membered ring. Preferably, it is saturated except for the double bond it shares with the dihydroquinazoline moiety. Preferably, it is unsubstituted. However, it may carry, for example, 1 or 2 substituents. Suitable substituents include halogen, hydroxyl, said C1-C4 alkyl such as methyl or ethyl, C1-C4 alkoxy such as methoxy or ethoxy, amino, (C1-C4 alkyl)amino, di-(C1-C4 alkyl)amino and xe2x80x94(C1-C4 alkyl)xe2x80x94OH.
A C2-C4 alkenyl group or moiety can be linear or branched but is preferably linear. Suitable such alkenyl groups and moieties include ethenyl, n-propenyl, i-propenyl and n-butenyl. Ethenyl and prop-2-enyl are preferred and prop-2-enyl is particularly preferred. A C2-C4 alkenyl group or moiety can be substituted or unsubstituted at any position. Typically, it carries up to three substituents, e.g. one or two substituents. Suitable substituents include hydroxy, halogen, amino, (C1-C4 alkyl)amino and di-(C1-C4 alkyl)amino.
A C2-C4 alkynyl group or moiety is an ethynyl, propynyl or butynyl group or moiety. Prop-2-ynyl is preferred. A C2-C4 alkynyl group or moiety can be substituted or unsubstituted at any position. Typically, it carries up to two substituients, e.g. one substituent. Suitable substituents include hydroxy, halogen, amino, (C1-C4 alkyl)amino and di-(C1-C4 alkyl)amino.
A divalent heteroaryl group is typically a 5- to 10-membered aryl ring containing at least one heteroatom selected from O, S and N. It may optionally be fused to a said aryl group or to a C3-C6 cycloalkyl group. It may be unsubsituted or substituted at any position. Typically, it carries up to three substituents, e.g. one or two substituents. Suitable substituents include halogen, hydroxyl, CF3, CCl3, said C1-C4 alkyl such as methyl or ethyl, C1-C4 alkoxy such as methoxy or ethoxy, amino, (C1-C4 alkyl)amino and di-(C1-C4 alkyl)amino.
Preferably, the divalent heteroaryl group is a 5- or 6-membered ring. More preferably, it is a divalent pyridyl, pyrimidinyl or thiazole group. Typically, it is attached to the rest of the molecule at each position via a carbon atom.
The 5- or 6-membered heterocyclic group may be an aromatic or non-aromatic, saturated or non-saturated, substituted or non-substituted such group. It may contain one or more, for example, one, two or three heteroatoms selected from N, O and S. Typically, it carries one, two or three substituents. Suitable substituents include halogen, hydroxy, CF3, CCl3, xe2x80x94CO2H, amino, (C1-C4 alkyl)amino and di(C1-C4alkyl)amino.
Suitable such heterocyclic groups include morpholinyl, imidazolidinyl, pyrazolidinyl, piperidyl and piperazinyl groups. Morpholinyl, for example morpholin-4-yl groups, and piperidyl groups are preferred.
The 5- or 6-membered N-containing heterocyclic ring may be an aromatic or non-aromatic, saturated or non-saturated, substituted or non-substituted ring. It may contain one or more, for example 1 or 2, further heteroatoms selected from N, O and S.
Typically, the N-containing heterocyclic ring carries up to three substituents, e.g. one or two substituents. Suitable substituents include halogen, hydroxyl, CF3, CCl3, said C1-C4 alkyl such as methyl, ethyl and hydroxyethyl, C1-C4 alkoxy such as methoxy or ethoxy, phenyl, -phenyl-CO2H, -phenyl-CO2xe2x80x94(C1-C4 alkyl), amino, (C1-C4 alkyl)amino, di-(C1-C4 alkyl)amino, xe2x80x94CO2H, xe2x80x94CO2xe2x80x94(C1-C4 alkyl) such as xe2x80x94CO2Me and a said 5- or 6-membered heterocyclic group such as a piperidyl, pyridyl, pyrazinyl, piperazinyl or morpholinyl group. Preferably, it is unsubstituted or substituted by phenyl, hydroxy, C1-C4 alkyl, for example methyl, ethyl or hydroxyethyl, or xe2x80x94CO2xe2x80x94(C1-C4 alkyl).
Suitable N-containing heterocyclic rings include piperazinyl, piperidinyl, morpholinyl and pyrrolidinyl rings.
When the compound of the invention contains two groups Rxe2x80x3, the two Rxe2x80x3 groups may be the same or different.
m is typically 1 or 2.
The moieties xe2x80x94(CH2)pxe2x80x94 and xe2x80x94(CH2)mxe2x80x94 are alkylene groups having p and m carbon atoms respectively. The alkylene groups may be substituted or unsubstituted. Typically, they carry 1 or 2 substituents. Suitable substituents include halogen, hydroxyl, CF3, CCl3, said C1-C4 alkyl such as methyl or ethyl, C1-C4 alkoxy such as methoxy or ethoxy, amino, (C1-C4 alkyl)amino and di-(C1-C4 alkyl)amino.
Preferably, R1 and R1xe2x80x2 together form an oxo group and R2 is as defined above or R1xe2x80x2 and R2 together form a bond and R1 is xe2x80x94SCH3, xe2x80x94NHCOPh or xe2x80x94NHRxe2x80x2 wherein Rxe2x80x2 is a C1-C4 alkyl group substituted by a di-(C1-C4 alkyl) amino group, for example xe2x80x94NHxe2x80x94CH2xe2x80x94CH2xe2x80x94N(CH3)2. More preferably, R1 and R1xe2x80x2 together form an oxo group and R2 is as defined above.
When R1 and R1xe2x80x2 together form an oxo group, R2 is preferably hydrogen, C1-C4 alkyl, for example methyl, ethyl or dimethylaminoethyl, xe2x80x94(C1-C4 alkyl)xe2x80x94CO2H, xe2x80x94(C1-C4alkyl)xe2x80x94CO2xe2x80x94(C1C4 alkyl), xe2x80x94(C1-C4 alkyl)xe2x80x94CONRxe2x80x2Rxe2x80x3 wherein Rxe2x80x2 and Rxe2x80x3 are the same or different and are selected from hydrogen and C1-C4 alkyl, xe2x80x94(C1-C2 alkyl)xe2x80x94B, xe2x80x94(C1-C2 alkyl)xe2x80x94COxe2x80x94(C1-C4 alkyl)xe2x80x94B, xe2x80x94(C1-C2 alkyl)xe2x80x94CO2xe2x80x94(C1-C4 alkyl)xe2x80x94B or xe2x80x94(C1-C2 alkyl)xe2x80x94CONHxe2x80x94(C1-C4 alkyl)xe2x80x94B, wherein B is as defined above, or xe2x80x94(C1-C2 alkyl)xe2x80x94COxe2x80x94Bxe2x80x2 wherein Bxe2x80x2 is a 5- or 6-membered heterocyclic group. More preferably, R2 is hydrogen, C1-C4 alkyl such as methyl, ethyl or dimethylaminoethyl, xe2x80x94(C1-C4 alkyl)xe2x80x94CO2H, xe2x80x94(C1-C4 alkyl)xe2x80x94CO2xe2x80x94(C1-C4 alkyl), xe2x80x94(C1-C4 alkyl)xe2x80x94CONRxe2x80x2Rxe2x80x3 wherein Rxe2x80x2 and Rxe2x80x3 are as defined above, or xe2x80x94(C1-C2 alkyl)xe2x80x94COBxe2x80x2 wherein Bxe2x80x2 is as defined above. Methyl, ethyl, xe2x80x94(CH2)2NMe2, xe2x80x94CH2xe2x80x94CO2H, xe2x80x94CH2xe2x80x94CO2Me, xe2x80x94CH2xe2x80x94COxe2x80x94NEt2 and xe2x80x94CH2xe2x80x94COxe2x80x94Nxe2x80x94 piperidyl are particularly preferred.
Preferably, R3 is xe2x80x94(CH2)pA wherein p is 1 or 2 and A is an optionally substituted piperidyl, piperazinyl, morpholinyl or pyrrolidinyl group or A is xe2x80x94NEt2. Suitable substituents include those given above as appropriate substituents for the N-containing heterocyclic ring. Preferred substituents are phenyl, hydroxy, C1-C4 alkyl, for example methyl, ethyl or hydroxyethyl, and xe2x80x94CO2xe2x80x94(C1-C4 alkyl).
More preferably, p is 1 and A is pyrrolidin-1-yl, methyl-L-prolin-1-yl, morpholin-4-yl, piperidin-1-yl, 4-hydroxy-piperidin-1-yl, 4-phenyl-piperazin-1-yl, 4-(2-hydroxyethyl)-piperazin-1-yl, piperazin-1-yl, 4-methylpiperazin-1-yl, 4-ethylpiperazin-1-yl or NEt2.
Typically, the 5- or 6-membered carbocyclic ring formed by R4 and R5 is a cyclopentene ring.
Preferably, R4 is hydrogen.
Preferably, R5 is halogen such as chlorine or bromine or is xe2x80x94CX3 wherein X is a halogen such as fluorine.
Preferably, Rxe2x80x3 is a C2-C4 alkenyl group or a C2-C4 alkynyl group.
Preferably, X1 is xe2x80x94NRxe2x80x3xe2x80x94 wherein Rxe2x80x3 is a C1-C4 alkyl, C2-C4 alkenyl or C2-C4 alkynyl group. More preferably, X1 is xe2x80x94N(CH3)xe2x80x94 or xe2x80x94N(CH2CCH)xe2x80x94.
When R4 and R5 do not together form a said carbocyclic ring, the moiety xe2x80x94CR4X1xe2x80x94 is typically xe2x80x94CON(C1-C4 alkyl)xe2x80x94 such as xe2x80x94CONMexe2x80x94, xe2x80x94CONHxe2x80x94, xe2x80x94CH(C1-C4 alkyl)NHxe2x80x94 such as xe2x80x94CHMeNHxe2x80x94, xe2x80x94CH(C1-C4 alkyl)N(C1-C4 alkyl)xe2x80x94 such as xe2x80x94CHMeNMexe2x80x94, xe2x80x94CH2Oxe2x80x94, xe2x80x94CH2Sxe2x80x94 or, preferably, xe2x80x94CH2N(CH2CCH)xe2x80x94. When R4 and R5, together with the carbon atoms to which they are attached, form a said carbocyclic ring, X1 is preferably xe2x80x94N(C1-C4 alkyl)xe2x80x94.
Preferably, Y is a phenylene or naphthylene group. More preferably, Y is a 1,4-phenylene group.
Preferably, R6 is oxo.
Preferably, X2 is xe2x80x94NHxe2x80x94 or xe2x80x94N(C1-C4 alkyl)xe2x80x94.
The moiety xe2x80x94CR6X2xe2x80x94 is typically xe2x80x94CH2N(CH2CCH)xe2x80x94, xe2x80x94CON(C1-C4 alkyl)xe2x80x94 such as xe2x80x94CONMexe2x80x94, xe2x80x94CH(C1-C4 alkyl)NHxe2x80x94 such as xe2x80x94CHMeNHxe2x80x94, xe2x80x94CH(C1-C4 alkyl)N(C1-C4 alkyl)xe2x80x94 such as xe2x80x94CHMeNMexe2x80x94, xe2x80x94CH2Oxe2x80x94, xe2x80x94CH2Sxe2x80x94 or, preferably, xe2x80x94COxe2x80x94NHxe2x80x94.
Preferably, R7 is pyridyl, imidazolyl, triazolyl, xe2x80x94(C1-C4 alkyl)-imidazolyl or xe2x80x94(C1-C4 alkyl)-triazolyl. More preferably, R7 is pyridyl, for example pyridin-3-yl, imidazolyl or triazolyl. As used herein, a triazolyl group is typically a 1,2,4-triazolyl or a 1,2,3-triazolyl group.
Preferred compounds of the invention are those in which:
either R1 and R1xe2x80x2 together form an oxo group and R2 is hydrogen, C1-C4 alkyl such as methyl, ethyl or dimethylaminoethyl, xe2x80x94(C1-C4 alkyl)xe2x80x94CO2H, xe2x80x94(C1-C4 alkyl)xe2x80x94CO2xe2x80x94(C1-C4alkyl), xe2x80x94(C1-C4 alkyl)xe2x80x94CONRxe2x80x2Rxe2x80x3 wherein Rxe2x80x2 and Rxe2x80x3 are as defined above, or xe2x80x94(C1-C2 alkyl)xe2x80x94COBxe2x80x2 wherein Bxe2x80x2 is as defined above, or R1xe2x80x2 and R2 together form a bond and R1 is xe2x80x94SCH3, xe2x80x94NHCOPh or xe2x80x94NHRxe2x80x2 wherein Rxe2x80x2 is a C1-C4 alkyl group substituted by a di-(C1-C4 alkyl)amino group, for example xe2x80x94NHxe2x80x94CH2xe2x80x94CH2xe2x80x94N(CH3)2;
R3 is xe2x80x94(CH2)pA wherein p is 1 or 2 and A is an optionally substituted piperidyl, piperazinyl, morpholinyl or pyrrolidinyl group or A is xe2x80x94NEt2;
R4 is hydrogen;
R5 is halogen such as chlorine or bromine or is xe2x80x94CX3 wherein X is a halogen such as fluorine;
the moiety xe2x80x94CR4X1xe2x80x94 is xe2x80x94CON(C1-C4 alkyl)xe2x80x94 such as xe2x80x94CONMexe2x80x94, xe2x80x94CONHxe2x80x94, xe2x80x94CH(C1-C4 alkyl)NHxe2x80x94 such as xe2x80x94CHMeNHxe2x80x94, xe2x80x94CH(C1-C4 alkyl)N(C1-C4 alkyl)xe2x80x94 such as xe2x80x94CHMeNMexe2x80x94, xe2x80x94CH2Oxe2x80x94, xe2x80x94CH2Sxe2x80x94 or, preferably, xe2x80x94CH2N(CH2CCH)xe2x80x94;
Y is a phenylene or naphthylene group;
the moiety xe2x80x94CR6X2xe2x80x94 is xe2x80x94CH2N(CH2CCH)xe2x80x94, xe2x80x94CON(C1-C4 alkyl)xe2x80x94 such as xe2x80x94CONMexe2x80x94, xe2x80x94CH(C1-C4 alkyl)NHxe2x80x94 such as xe2x80x94CHMeNHxe2x80x94, xe2x80x94CH(C1-C4 alkyl)N(C1-C4 alkyl)xe2x80x94 such as xe2x80x94CHMeNMexe2x80x94, xe2x80x94CH2Oxe2x80x94, xe2x80x94CH2Sxe2x80x94 or, preferably, xe2x80x94COxe2x80x94NHxe2x80x94;
m is 1; and
R7 is pyridinyl, for example pyridin-3-yl, imidazolyl or triazolyl
and pharmaceutically acceptable salts thereof
Typically, in the above preferred compounds, when R1 and R1xe2x80x2 together form an oxo group, R2 is C1-C4 alkyl such as methyl. Typically, in the above preferred compounds, R7 is 3-pyridyl.
Further preferred compounds of the invention are compounds of the formula (Ia) and pharmaceutically acceptable salts thereof, 
wherein:
R2 is hydrogen, C1-C4 alkyl such as methyl, ethyl or dimethylarminoethyl, xe2x80x94(C1-C4 alkyl)xe2x80x94CO2H, xe2x80x94(CC4 alkyl)xe2x80x94CO2xe2x80x94(C1-C4 alkyl), xe2x80x94(C1-C4 alkyl)xe2x80x94CONRxe2x80x2Rxe2x80x3 wherein Rxe2x80x2 and Rxe2x80x3 are as defined above, or xe2x80x94(C1-C2 alkyl)xe2x80x94COBxe2x80x2 wherein Bxe2x80x2 is as defined above;.
R3 is xe2x80x94(CH2)pxe2x80x94A wherein p is from 1 to 4 and A is a 5- or 6-membered N-containing heterocyclic ring or A is xe2x80x94NAxe2x80x2Axe2x80x3 wherein Axe2x80x2 and Axe2x80x3 are the same or different and are both C1-C4 alkyl groups;
R5 is chlorine or bromine; and
R7 is pyridyl, for example pyridin-3-yl, imidazolyl or triazolyl.
R2 in the formula (Ia) can be hydrogen or C1-C4 alkyl. It is preferably methyl, ethyl, xe2x80x94(CH2)2NMe2, xe2x80x94CH2xe2x80x94CO2H, xe2x80x94CH2xe2x80x94CO2Me, xe2x80x94CH2xe2x80x94COxe2x80x94NEt2 or xe2x80x94CH2xe2x80x94COxe2x80x94N-piperidinyl. R3 in the formula (Ia) is preferably xe2x80x94(CH2)pA wherein p is 1 or 2 and A is an optionally substituted piperidyl, piperazinyl, morpholinyl or pyrrolidinyl group or A is xe2x80x94NEt2. More preferably, p is 1 and A is pyrrolidin-1-yl, methyl-L-prolin-1-yl, morpholin-4-yl, piperidin-1-yl, 4-hydroxy-piperidin-1-yl, 4-phenyl-piperazin-1-yl, 4-(2-hydroxyethyl)-piperazin-1-yl, piperazin-1-yl, 4-methylpiperazin-1-yl, 4-ethylpiperazin-1-yl or xe2x80x94NEt2. R7 in the formula (Ia) is preferablypyridin-3-yl.
Further preferred compounds of the invention are compounds of formula (Ib) 
wherein R2, R3 and R7 are as defined in the formula (Ia) and Rxe2x80x3 is hydrogen, C1-C4 alkyl, C2-C4 alkenyl or C2-C4 alkynyl. Preferably, Rxe2x80x3 in the formula (Ib) is C1-C4 alkyl, for example methyl.
The present invention includes pharmaceutically acceptable salts of the compounds of formula (I) and of formula (Ia). Suitable salts include salts with pharmaceutically acceptable acids, both inorganic acids such as hydrochloric, sulphuric, phosphoric, diphosphoric, hydrobromic or nitric acid and organic acids such as citric, fumaric, maleic, malic, ascorbic, succininc, tartaric, benzoic, acetic, methanesulphonic, ethanesulphonic, benzenesulphonic or p-toluenesulphonic acid. Salts may also be formed with pharmaceutically acceptable bases such as alkali metal (eg sodium or potassium) and alkali earth metal (eg calcium or magnesium) hydroxides and organic bases such as alkyl amines, aralkyl amines or heterocyclic amines. A preferred salt is the hydrochloride salt.
Some of the compounds of the invention include one or more chiral centre. The present invention includes enantiomers and diastereoisomers of such compounds. For example, the group X2xe2x80x94(CH2)mxe2x80x94R7 may include a chiral centre when the alkylene moiety xe2x80x94(CH2)mxe2x80x94 is substituted.
Particularly preferred compounds of the invention are:
4-[N-[7-chloro-3-methyl-2-(4-methyl-piperazin-1-yl)methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide,
4-[N-[2-diethylaminomethyl-7-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide,
4-[N-[7-chloro-3-methyl-4-oxo-2-(piperldin-1-yl)methyl-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide,
4-[N-[7-chloro-3-methyl-2-(morpholin-4-yl)methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide,
4-[N-[7-chloro-3-methyl-2-(pyrrolidin-1-yl)methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide,
4-[N-[7-chloro-3-methyl-2-(4-ethyl-piperazin-1-yl)methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide,
4-[N-[7-chloro-3-methyl-2-(methyl-L-prolin-1-yl)methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide,
4-[N-[7-chloro-3-methyl-2-(4-(2-hydroxyethyl)-piperazin-1-yl)methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide,
4-[N-[7-chloro-3-methyl-4-oxo-2-(4-phenylpiperazin-1-yl)methyl-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide,
4-[N-[7-chloro-3-methyl-2-(4-hydroxypiperidin-1-yl)methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide,
4-[N-[7-chloro-3-methyl-2-(4-methyl-piperazin-1-yl)methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-[3-(1H-imidazol-1-yl)propyl)benzamide,
4-[N-[7-chloro-3-methyl-4-oxo-2-(piperidin-1-yl)methyl-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-[(3-(1H-1,2,4-triazol-1-yl)propyl]benzamide,
4-[N-[7-chloro-3-diethylcarbamoylmethyl-4-oxo-2-(piperidin-1-yl)methyl-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide,
4-[N-[7-chloro-4-oxo-2-(piperidin-1-yl)methyl-3-piperidinocarbonylmethyl-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide,
4-[N-[7-chloro-3-methoxycarbonylmethyl-4-oxo-2-piperidin-1-ylmethyl-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide,
4-[N-[7-chloro-3-(2-dimethylaminoethyl)-4-oxo-2-piperidin-1-ylmethyl-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide,
4-[N-[3-methyl-4-oxo-2-(piperidin-1-yl)methyl-3,4,7,8-tetrahydro-6H-cyclopenta[g]quinazolin-6-yl]-N-methylamino]-N-(3-pyridylmethyl)benzamide
and pharmaceutically acceptable salts thereof.
Compounds of the invention may be prepared by reacting a compound of formula (III)
X2xe2x80x2xe2x80x94(CH2)mxe2x80x94R7xe2x80x83xe2x80x83(III)
wherein m and R7 are as defined above and X2xe2x80x2 is OH, SH or NH2Rxe2x80x3 wherein
Rxe2x80x3 is as defined above, with a compound of formula (IIa) or (IIb) 
wherein R1, R1xe2x80x2, R2, R3, R4, R5, X1 and Y are as defined above, R6 is hydrogen or C1-C4 alkyl and L is a leaving group such as bromine.
The reaction between the compounds of formula (IIa) and (III) typically involves two stages. In the first stage, the tertiary butyl ester is hydrolysed. The hydrolysis is typically carried out in a solvent such as dichloromethane, in the presence of an acid such as trifluoroacetic acid at a temperature from 0xc2x0 C. to the reflux temperature of the solvent. Typically, it is conducted at room temperature.
In the second stage, the compound of formula (III) is typically reacted with the hydrolysed pro duct in a dipolar aprotic solvent such as anhydrous DMF, using a coupling agent such as PyBOP(copyright). Typically, the reaction takes place in the presence of a base such as diisopropylethylamine. Alternatively, the hydrolysed product may be converted to an acyl chloride, for example by reaction with thionyl chloride, and the compound of formula (III) may be reacted with the acyl chloride.
The leaving group L is preferably a halogen such as bromine. Typically, the reaction between the compounds of formula (IIb) and (III) takes place in a polar aprotic. solvent such as anhydrous DMF at a temperature from 0xc2x0 C. to the reflux temperature of the solvent. Preferably, it is conducted in the presence of a base such as 2,6-lutidine. The reaction typically takes place under an inert atmosphere such as an argon atmosphere.
A compound of formula (IIb) may be prepared from a corresponding compound of formula (IIb) in which L is a hydrogen atom. For example, a compound of formula (IIb) in which L is a hydrogen atom can be reacted with N-bromosuccinimide in a solvent such as anhydrous carbon tetrachloride. Typically, such a reaction is conducted in the presence of an initiator such as dibenzoyl peroxide under strong illumination,
The compounds of formula (III) are commercially available known compounds or may be made by analogy with known methods.
Compounds of formula (IIa) or (IIb) in which R1 is xe2x80x94Sxe2x80x94(C1-C4 alkyl) can be prepared from corresponding compounds in which R1 and R1xe2x80x2 together form an oxo group and in which R2 is hydrogen.
To effect the conversion, the latter compounds may be reacted with Lawesson""s Reagent (2,4-bis-(4-methoxyphenyl)-1,3-dithio-2,4-diphosphetane-2,4-disulfide) in a solvent such as toluene at, for example, the reflux temperature of the solvent. The 4-oxo group can be thereby converted to a 4-thioxo group. The 4-thioxo compounds may be reacted with (C1-C4 alkyl)xe2x80x94I in the presence of a base such as Cs2 CO3 in a solvent such as DMF to give compounds in which R1 is xe2x80x94Sxe2x80x94(C1-C4 alkyl).
Compounds of formula (IIa) or (IIb) in which R1 is xe2x80x94NHRxe2x80x2 or xe2x80x94NHCORxe2x80x2 in which Rxe2x80x2 is as defined above may be prepared from corresponding compounds in which R1 is xe2x80x94Sxe2x80x94(C1-C4 alkyl) by reacting the latter compounds with H2NRxe2x80x2 or H2NCORxe2x80x2 wherein Rxe2x80x2 is as defined above in a solvent such as DMF at a temperature above room temperature, for example at the reflux temperature of the solvent.
The compounds of formula (IIa) and compounds of formula (IIb) in which L is a hydrogen atom can be prepared by reacting a compound of formula (IV) 
wherein R1, R1xe2x80x2, R2, p, R4, R5, X1 and Y are as defined above, Z is xe2x80x94CO2xe2x80x2Bu or xe2x80x94CH2R6 wherein R6 is hydrogen or C1-C4 alkyl and L is a leaving group such as a methanesulphonyloxy group (mesylate group), a tosylate group, a triflate group or a halogen such as bromine, with a group AH wherein A is as defined above. Typically, the reaction is conducted in a solvent such as anhydrous dichloromethane under an inert atmosphere such as an argon atmosphere.
The compounds of formula (IV) can be prepared by reacting a compound of formula (V) 
wherein R1, R1xe2x80x2, R2, p, R4, R5, X1, Y and Z are as defined above, with a reagent such as methanesulphonic anhydride, triflic anhydride, tosyl anhydride or PX3 wherein X is a halogen such as bromine. Typically, the reaction takes place in a solvent such as anhydrous dichloromethane in the presence of a base such as triethylamine. Typically, the reaction takes place at below room temperature, for example at around 0xc2x0 C.
The compounds of formula (V) can be made by hydrolysing an ester of formula (VI) 
wherein R1, R1xe2x80x2, R2, p, R4, R5, X1, Y and Z are as defined above.
Typically, the hydrolysis is effected by reaction with aqueous NaOH and water in the presence of a solvent such as tetrahydrofuran.
The esters of formula (VI) can be prepared by reacting a compound of formula (VII) 
wherein p, R1, R1xe2x80x2, R2, R4 and R5 are as defined above and L is a leaving group such as bromine, with a compound of formula (VIII)
X1xe2x80x2xe2x80x94Yxe2x80x94Zxe2x80x83xe2x80x83(VIII)
wherein X1xe2x80x2 is OH, SH or xe2x80x94NHRxe2x80x3 wherein Rxe2x80x3 is as defined above and Y and Z are as defined above.
Typically, the reaction is carried out in a polar aprotic solvent such as anhydrous DMF at a temperature from 0xc2x0 C. to the reflux temperature of the solvent. Preferably, the reaction takes place at around 120xc2x0 C. Preferably, it is conducted in the presence of a base such as 2,6-lutidine. The reaction typically takes place under an inert atmosphere such as an argon atmosphere. The compound of formula (VIII) may be reacted with NaH to increase its nucleophilicity before it is reacted with the compound of formula (VII).
Compounds of formula (VI) in which R4 and R5 together form a carbocyclic ring and in which X1 is xe2x80x94NRxe2x80x3xe2x80x94 wherein Rxe2x80x3 is as defined above, can be prepared by reductive amination of a compound of formula (VIIxe2x80x2) 
wherein R1, R1xe2x80x2, R2, and p are as defined above and a is 1 or 2. Such reductive amination can be effected by reacting a compound of formula (VIIxe2x80x2) with a said compound of formula (VIII) in which X1xe2x80x2 is xe2x80x94NHRxe2x80x3 wherein Rxe2x80x3 is as defined above. Typically, the reaction takes place in three stages.
In the first stage, a compound of formula (VIIxe2x80x2) and a compound of formula NH2xe2x80x94Yxe2x80x94Z, wherein Y and Z are as defined above, are typically reacted under acidic conditions, for example in the presence of p-toluenesulphonic acid, in a solvent such as 1,2-dimethoxyethane at the reflux temperature of the solvent.
In the second stage, the thus obtained imine is typically reduced with a reducing agent such as NaCNBH3 in the presence of an acid such as AcOH in a solvent such as methanol.
In the third stage, the thus obtained amine is typically reacted with an aldehyde of formula Rxe2x80x3xe2x80x94COH wherein Rxe2x80x3 is as defined above, in the presence of an acid such as AcOH in a solvent such as THF, and then reduced with a reducing agent such as NaCNBH3.
The compounds of formula (VIIxe2x80x2) can be prepared by oxidising a compound of formula 
in which R1, R1xe2x80x2, R2, p and a are as defined above. The oxidation can be effected, for example, with a 70% t-butylhydroperoxide solution in the presence of chromium (VI) oxide in a solvent such as CH2Cl2.
The compounds of formula (VIII) are commercially available known compounds or may be made by analogy with known methods.
The compounds of formula (VII) in which R4 is other than oxo can be prepared by reacting a corresponding compound of formula (VII) in which L is a hydrogen atom with a reagent such as N-bromosuccinimide. Typically, the reaction takes place in a solvent such as anhydrous carbon tetrachloride under an inert atmosphere such as an argon atmosphere in the presence of an initiator such as dibenzoyl peroxide and under strong illumination. The compounds of formula (VII) in which R4 is oko can be prepared by reacting a corresponding compound of formula (VII) in which L is a hydroxy group with a reagent such as oxalyl chloride or thionyl chloride.
Compounds of formula (IX) 
wherein R1, R1xe2x80x2, R2, p and R5 are as defined above and Zxe2x80x2 is xe2x80x94CO2H or xe2x80x94CH2R4 in which R4 is other than oxo, can be prepared by reacting a compound of formula (X) 
wherein R1, R1xe2x80x2, R2, p, R5 and Zxe2x80x2 are as defined above and L is a leaving group such as chlorine, with caesium acetate. Typically, the reaction takes place in a solvent such as anhydrous DMF under an inert atmosphere such as an argon atmosphere at a temperature of from 0xc2x0 C. to the reflux temperature of the solvent, preferably around 85xc2x0 C.
Compounds of formula (IX) or (X) in which R2 is C1-C4 alkyl, xe2x80x94(C1-C4 alkyl)xe2x80x94COB, xe2x80x94(C1-C4 alkyl)xe2x80x94COxe2x80x94(C1-C4 alkyl)xe2x80x94B, xe2x80x94(C1-C4 alkyl)xe2x80x94CO2xe2x80x94(C1-C4 alkyl)xe2x80x94B, xe2x80x94(C1-C4 alkyl)xe2x80x94CO2xe2x80x94(C2-C4 alkenyl)xe2x80x94B or xe2x80x94(C1-C4 alkyl)xe2x80x94CONHxe2x80x94(C1-C4 alkyl)xe2x80x94B wherein B is as defined above, can be prepared from a corresponding compound in which R2 is hydrogen by reaction with R2xe2x80x94I wherein R2 is C1-C4 alkyl, xe2x80x94(C1-C4 alkyl)xe2x80x94COB, xe2x80x94(C1-C4 alkyl)xe2x80x94COxe2x80x94(C1-C4 alkyl)xe2x80x94B, xe2x80x94(C1-C4 alkyl)xe2x80x94CO2xe2x80x94(C1-C4 alkyl)xe2x80x94B, xe2x80x94(C1-C4 alkyl)xe2x80x94CO2xe2x80x94(C2-C4 alkenyl)xe2x80x94B or xe2x80x94(C1-C4 alkyl)xe2x80x94CONHxe2x80x94((C1-C4 alkyl)xe2x80x94B wherein B is as defined above. Typically, the reaction takes place in the presence of a base such as sodium hydride under an inert atmosphere such as an argon atmosphere.
The compounds of formula (X) can be made by analogy with known methods. For example, compounds of formula (X) in which R1 and R1xe2x80x2 together form an oxo group can be prepared by reacting a compound of formula (XI) 
wherein Zxe2x80x2 and R5 are as defined above, with CH2ClCN (chloroacetonitrile). Typically, the reaction takes place in a solvent such as anhydrous methanol in the presence of a base such as sodium methoxide at room temperature under an inert atmosphere such as an argon atmosphere.
The substituted benzene compounds of formula (XI) can be prepared, for example, by reacting a 2-cyanoaniline compound substituted at the 4-position with Zxe2x80x2 and at the 5-position with R5, with hydrogen peroxide. The said 2-cyanoaniline compound can be prepared from a corresponding 2-bromoaniline compound by reaction with Cu(I) cyanide. The said 2-bromoaniline compound can be prepared by reacting a corresponding aniline compound unsubstituted at the 2-position with bromine in a diethyl ether/acetic acid solvent at below 5xc2x0 C. under an inert atmosphere.
The compounds of the invention have anti-cancer activity. They are active against leukaemias, lymphoid malignancies and solid tumours such as carcinomas and sarcomas. They can therefore be used to treat leukaemia, breast cancer, lung cancer, livercancer, cancer of the colon, rectal cancer, stomach -cancer, prostate cancer, cancer of the bladder, pancreatic cancer and ovarian cancer.
The compounds of the present invention have a different pattern of activity to known chemotherapeutic agents. It is therefore thought that the compounds of the invention act via a new, non-folate dependent, locus. The locus at which the compounds of the invention act is thought to be different from the locii targeted by known chetnotlierapcutic agents.
The present invention includes a pharmaceutical composition comprising a compound according to the invention and a pharmaceutically acceptable carrier or diluent.
The compounds of the invention may be administered in a variety of dosage forms. Thus, they can be administered orally, for example as tablets, troches, lozenges, aqueous or oily suspensions, dispersiblepowders or granules. The compounds of the invention may also be administered parenterally, either subcutaneously, intravenously, intramuscularly, intrasternally, transdermally or by infusion techniques. The compounds may also be administered as suppositories.
A compound of the invention is typically formulated for administration with a pharmaceutically acceptable carrier or diluent. For example, solid oral forms may contain, together with the active compound, diluents, e.g. lactose, dextrose, saccharose, cellulose, corn starch or potato starch; lubricants, e.g. silica, talc, stearic acid, magnesium or calcium stearate, and/or polyethylene glycols; binding agents; e.g. starches, arabic gums, gelatin, methylcellulose, carboxymethylcellulose or polyvinyl pyrrolidone; disaggregating agents, e.g. starch, alginic acid, alginates or sodium starch glycolate; effervescing mixtures; dyestuffs; sweeteners; wetting agents, such as lecithin, polysorbates, laurylsulphates; and, in general, non-toxic and pharmacologically inactive substances used in pharmaceutical formulations. Such pharmaceutical preparations may be manufactured in known manner, for example, by means of mixing, granulating, tabletting, sugar-coating, or film coating processes.
Liquid dispersions for oral administration may be syrups, emulsions and suspensions. The syrups may contain as carriers, for example, saccharose or saccharose with glycerine and/or mannitol and/or sorbitol.
Suspensions and emulsions may contain as carrier, for example a natural gum, agar, sodium alginte, pectin, methylcellulose, carboxymethylcellulose, or polyvinyl alcohol. The suspensions or solutions for intramuscular injections may contain, together with the active compound, a pharmaceutically acceptable carrier, e.g. sterile water, olive oil, ethyl oleate, glycols, e.g. propylene glycol, and if desired, a suitable amount of lidocaine hydrochloride.
Solutions for intravenous or infusions may contain as carrier, for example, sterile water or preferably they may be in the form of sterile, aqueous, isotonic saline solutions.
A therapeutically effective amount of a compound of the invention is administered to a patient. A typical daily dose is up to 50 mg per kg of body weight, for example from 0.001 to 50 mg per kg of body weight, according to the activity of the specific compound, the age, weight and conditions of the subject to be treated, the type and severity of the disease and the frequency and route of administration. Preferably, daily dosage levels are from 0.05 mg to 2 g, preferably from 0.1 mg to 10 mg.
The following Examples illustrate the invention.
To a solution of 3-chloro-4-methylaniline (10.0 g, 70.6 mmol) in diethyl ether/acetic acid (v/v 1/1, 350 ml) cooled in an ice-bath was dropwise added bromine (4 ml) over a 35 min period under an argon atmosphere while the temperature of the reaction mixture was kept below 5xc2x0 C. Stirring was continued for 10 min after the addition of bromine; then the yellow reaction mixture was partitioned between dichloromethane (250 ml) and dilute brine (200 ml). The organic layer was washed with more dilute brine (200 ml), dried (Na2SO4), and concentrated in vacuo to an oily residue. This was redissolved in dichloromethane (200 ml) and the solution was washed with saturated aqueous sodium bicarbonate (3xc3x97200 ml; caution: gas is evolved), dried (Na2SO4) and concentrated in vacuo to leave a brown wet solid. Purification by column chromatography on gradient elution with dichloromethane in hexanes (25 to 30%) gave in order of elution:
a. 5-chloro-2,6-dibromo-4-methylaniline as a white solid (4.63 g) mp 77-78xc2x0 C.
b. the desired product, 2-bromo-5-chloro-4-methylaniline (5.32 g, 34%), mp 90xc2x0 C., 1H-NMR (CDCl3) 2.23 (s, 3H, 4-CH3), 3.99 (br s, 2H, NH2), 6.78 (s, 1H, 6-H), 7.26 (s, 1H, 3-H). MS (FAB, m/z) 219, 221, 223 [(M+H)+, BrCl isotopic pattern]. Elemental Analysis: Found: C, 38.04; H, 3.20; N, 6.35; Cl, 16.04; Br, 36.28. C7H7BrClN requires: C, 38.13; H, 3.20; N, 6.35; Cl, 16.08; Br, 36.24
c. 2-bromo-3-chloro-4-methylaniline as a white solid (1.22 g) mp 48-56xc2x0 C.
Acetic anhydride (78 ml, 0.825 mol) was added in portions (using a dropping funnel) during a 45 min period to a stirred, cooled in an ice-bath solution of 3-chloro-4-methylaniline (106.2 g, 0.75 mol) in ethyl acetate (550 ml, dried over MgSO4 prior to use) and anhydrous pyridine (66.6 ml, 0.825 mol) under argon. During the reaction the temperature of the reaction mixture varied between 10-20xc2x0 C. Stirring was continued for 20 min; then the ice-water bath was removed and the reaction mixture was stirred for 18 h at room temperature. The solvents were removed in vacuo and the light brown solid residue was triturated with ether (350 ml) and left to stand in a fridge overnight. The solid was collected by filtration, washed with cold ether (100 ml) and hexanes (100 mL), dried over P2O5 to afford a white solid (90 g). The filtrate was concentrated in vacuo, triturated with ether to afford an additional 28.1 g of the product. Total yield 118.1 g (86%); mp 105-106xc2x0 C.; 1H-NMR (CDCl3) 2.17, 2.32 (2xc3x97s, 6H, 4-CH3, CH3CO), 7.14 (d, J=8.22 Hz, 1H, 6-H), 7.25 (2xc3x97dd, J=1.9, 6.5 Hz, 1H, 5-H), 7.58 (d, J=1.8 Hz, 1H, 3-H); MS (FAB, m/z) 184, 186 [(M+H)+ 100%, 30% respectively; Cl isotopic pattern].
Elemental analysis: Found C, 58.76; H, 5.46; N, 7.59; Cl, 19.20. C9H10ClNO requires: C, 58.87; H, 5.49; N, 7.63; Cl, 19.30%.
To a solution of 4-acetamido-2-chlorotoluene (89.2 g, 0.486 mol) in glacial acetic acid (480 ml) that was stirred with an overhead mechanical stirrer and under argon was dropwise added bromine (28.5 ml) during a period of 2 h while the temperature of the reaction mixture was kept below 15xc2x0 C. by using an ice-bath. Stirring was continued for a further 1.5 h after the addition of bromine, under an argon atmosphere. Then the brownish reaction mixture was poured into ice-water (1.8 lt), with the aid of water (1 lt), washed with water (6 lt), and dried in vacuo over P2O5. Purification by recrystallisation from acetonitrile afforded the desired product as a white crystals (61.5 g, 48%), mp 154-155xc2x0 C.; 1H-NMR (CDCl3) 2.24, 2.32 (2xc3x97s, 6H, 2xc3x97CH3), 7.40, 7.73 (2xc3x97s, 2H, 3-H, 6-H), 7.49 (br s, 1H, CONH); MS (FAB, m/z): 262, 264, 266 [(M+H)+, 80%, 100%, 25% respectively; BrCl isotopic pattern];
Elemental analysis, Found C, 41.17; H, 3.44; N, 5.36; Cl, 13.44, Br 30.60. C9H9BrClNO requires C, 41.17; H, 3.46; N, 5.34; Cl, 13.50; Br, 30.44.
A solution of 4-acetamido-5-bromo-2-chlorotoluene (64 g, 0.245 mol) in glacial acetic acid (48 ml) and concentrated hydrochloric acid (96 ml) was heated at 118xc2x0 C. for 24 h. The reaction mixture was allowed to cool to room temperature, diluted with water (200 ml), cooled in an ice-bath and the pH was. adjusted to 5 with a aqueous solution of NaOH (50% w/v). The precipitate was collected by filtration washed with water, and dried in vacuo over P2O5 to afford a white solid (50.7 g, 94%), mp 90xc2x0 C.
To a solution of 2-bromo-5-chloro-4-methylaniline (13.0 g 58.96 mmol) in N-methylpyrrolidinone (100 ml) was added copper(I) cyanide (10.56 g, 118 mmol). The reaction mixture was placed in an oil bath preheated to 163xc2x0 C. and stirred at this temperature for 2 h. The reaction mixture was allowed to cool to room temperature and then poured in ice-water (300 ml) and aqueous ammonia (90 ml). The brown precipitate was collected by filtration, washed with water (150 ml), dissolved in dichloromethane and the insoluble material was removed by filtration. The filtrate was washed with brine (100 ml), dried (MgSO4) and concentrated in vacuo. Purification, on gradient elution with dichloromethane in petroleum ether 60-80xc2x0 C. (65 to 95%), afforded a white solid (6.52 g), mp 180xc2x0 C.; 1H-NMR (DMSO-d6) 2.14 (s, 3H, 4-CH3), 6.11 (s, 2H, NH2), 6.86 (s, 1H, 6-H), 7.39 (s, 1H, 3-H); MS (FAB, m/z) 166,168 [(M+H)+; 90%, 40% respectively; Cl isotopic pattern]; FAB-HRMS: measured 166.0307; calculated for C8H8ClN2 (M+H)+ 166.0298.
A mixture of 5-chloro-2-cyano-4-methylaniline (4.0 g,0.024 mol), 30% aqueous KOH solution (56 ml), and 30% hydrogen peroxide (4 ml) was placed in an oil bath preheated to 130xc2x0 C., then stirred at this temperature for 2 hours (a clear solution had obtained after 1.5 h). The clear solution was then allowed to cool to room temperature, diluted with water (200 ml), and acidified to pH xcx9c5.50 with 3N HCl, then allowed to stand at room temperature for few hours. The off white solid was collected by filtration, washed with water and dried in vacuo over P2O5 (4.13 g, 93%), mp 212-215xc2x0 C. 1H-NMR (DMSO-d6) 2.16 (s, 3H, CH3), 6.83, 7.62 (2xc3x97s, 2H, 3-H, 6-H), 8.50 (br s, 2H, NH2); MS (FAB, m/z): 188, 186 (M+H)+; FAB-HRMS: measured: 185.0256; calculated for C8H8ClNO2 (M+) 185.0244.
To a flask containing sodium (36 mg) was added anhydrous MeOH (5 ml). Chloroacetonitrile (0.520 g, 6.9 mmol) was then added and the clear solution was stirred at room temperature for 30 min under argon. A solution of 2-amino-4-chloro-5-methylbenzoic acid (1.13 g, 6.0 mmol) in anhydrous methanol (25 ml) was then added with a syringe via a rubber septum. The reaction mixture was stirred at room temperature for 2 hours under argon, then it was fitted with a condenser and placed in an oil bath preheated to 80xc2x0 C. Stirring was continued at this temperature for 2 hours under argon, then the reaction mixture was allowed to cool to room temperature. The precipitate was collected by filtration, washed with methanol (10 ml), and water (10 ml), and dried in vacuo over P2O5 to afford a grey solid (1.0 g, 69%), mp 287-290xc2x0 C.; 1H-NMR (DMSO-d6) 2.47 (s, 3H, 6-CH3), 4.53 (s, 2H, CH2Cl), 7.75, 8.08 (2xc3x97s, 2H, 5-H and 8-H); 12.60 (s, 1H, N3xe2x80x94H); MS (FAB, m/z) 243, 244, 245 (M+H)+.
Elemental Analysis: Found: C, 49.22; H, 3.40; N, 11.39; Cl, 28. 94. C10H8Cl2N2O requires C, 49.41; H, 3.32; N, 11.52; Cl, 29.17%.
A mixture of 7-chloro-2-chloromethyl-6-methyl-3,4-dihydroquinazolin-4-one (0.500 g, 2.06 mmol), anhydrous DMF (14 ml) and cesium acetate (1.58 g, 8.24 mmol) was placed in an oil bath preheated to 85xc2x0 C., then stirred at this temperature for 2 hours and 15 min under argon. The reaction mixture was then allowed to cool to room temperature and the solvent was removed in vacuo. The residue was treated with hexanes (20 ml), collected by filtration, washed with hexanes (20 ml), water, and dried in vacuo over P2O5 (0.476 g, 87%), mp 220-225xc2x0 C., 1H-NMR (DMSO-d6) 2.14 (s, 3H, CH3CO), 2.45 (s, 3H, 6-CH3), 4.95 (s, 2H, 2-CH2O), 7.70, 8.06 (2xc3x97s, 2H, 5-H and 8-H), 12.44 (s, 1H, N3xe2x80x94H).
MS (FAB, m/z) 267 (M+H)+. FAB-HRMS measured 267.0520, calculated for C12H12ClN2O3 (M+H)+ 267.0536.
To a suspension of 2-acetoxymethyl-7-chloro-6-methyl-3,4-dihydroquinazolin-4-one 0.428 g, 1.6 mmol) in anhydrous DMF (13 ml) was added sodium hydride (60% dispersion, 0.070 g, 1.76 mmol) under argon. Stirring was continued at room temperature for 1 min and then iodomethane (0.20 ml, 3.2 mmol) was added into the reaction mixture with a syringe via a septum. Stirring was continued at room temperature for 1 hour and the reaction mixture was then partitioned between ethyl acetate (130 ml) and brine (80 ml). The organic layer was washed with dilute brine (80 ml), dried (Na2SO4) and concentrated in vacuo to leave an orange residue. This orange residue was dissolved in dichloromethane and to this solution silica gel (Art 7734, 1.7 g) was added. The solvent was removed in vacuo and the orange free running powder was placed on a silica gel column made up in 5% ethyl acetate in dichloromethane. The column was eluted with 5% ethyl acetate in dichloromethane to afford a pale yellow solid (0.300 g, 67%), mp 110-112xc2x0 C.; 1H-NMR (DMSO-d6) 2.17 (s, 3H, CH3CO), 2.47 (s, 3H, 6-CH3), 3.54 (s, 3H, N3xe2x80x94CH3), 5.23 (s, 2H, 2-CH2O), 7.71, 8.09 (2xc3x97s, 2H, 5-H, 8-H).
MS (FAB, m/z) 281,283 (M+H)+, 100%, 25%; Cl isotopic pattern) Elemental Analysis: Found: C, 55.57; H, 4.70; N, 10.01; Cl, 12.67. C13H13ClN2O3 requires:C, 55.62; H, 4.67; N, 9.98; Cl, 12.63%.
To a nearly clear solution of 2-acetoxymethyl-7-chloro-3,6-dimethyl-3,4-dihydroquinazolin-4-one (2.85 g, 10.16 mmol) in anhydrous carbon tetrachloride (60 ml) under argon was added N-bromosuccinimide (1.99 g, 11.17 mmol) followed by dibenzoyl peroxide (25 mg). The reaction flask was then fitted with a condenser and placed in an oil bath preheated to 85xc2x0 C., and illuminated with two 60 W bulbs. Stirring was continued at this temperature for 3 hours and 50 min, then the reaction mixture was allowed to cool to room temperature. The white precipitate was filtered off, washed with dichloromethane and the filtrate was concentrated in vacuo to leave a white solid. This was partitioned between ethyl acetate (200 ml)/dichloromethanre (25 ml) and dilute brine (100 ml). The organic layer was washed with more dilute brine (100 ml), dried (Na2SO4) and concentrated in vacuo. The white solid was dissolved in dichloromethane and to this solution silica gel (Art 7734, 3.5 g) was added. The solvent was removed in vacuo and the white free running powder was placed on a silica gel column made up in 5% ethyl acetate in dichloromethane. The column was eluted with a gradient of ethyl acetate in dichloromethane (5 to 10%). Pure by TLC fractions were combined, concentrated in vacuo to leave a white solid which was triturated with ethyl acetate/hexanes (v/v 4:6, 20 ml). The white solid was collected by filtration, and dried in vacuo (1.83 g, 51%), mp 183-186xc2x0 C. 1H-NMR (DMSO-d6) 2.18 (s, 3H, COCH3), 3.52 (s, 3H, N3-Me), 4.92 (s, 2H, CH2Br), 5.25 (s, 2H, 2-CH2O), 7.78, 8.39 (2xc3x97s, 2H, 5-H, 8-H).
MS (FAB, m/z): 363,361,359 [(M+H)+, 30%, 100%, 80% respectively; BrCl isotopic pattern]. Elemental analysis: Found: C, 43.62; H, 3.36; N, 7.77; Br, 9.73; Cl, 21.91. C13H12BrClN2O3 requires: C, 43.42; H, 3.36; N, 7.79; Br, 9.86; Cl, 22.22.
A flask containing 2-acetoxymethyl-6-bromomethyl-7-chloro-3-methyl-3,4-dihydroquinazolin-4-one (1.75 g, 4.88 mmol), anhydrous DMF (30 ml), tert-butyl 4-N-(prop-2-ynyl)aminobenzoate (1.35 g, 5.86 mmol), and 2,6-lutidine (1.38 g, 12.89 mmol) was fitted with a condenser, and placed in an oil bath preheated to 120xc2x0 C., then stirred at this temperature for 5.5 hours under argon. The solvent was next removed in vacuo and the brown residue was partitioned between ethyl acetate (350 ml) and brine (120 ml). The organic layer was washed with more dilute brine (120 ml), dried (Na2SO4) and concentrated in vacuo. The brown residue was purified by column chromatography using dichloromethane:ethyl acetate: petroleumn ether 60-80xc2x0 C. (v/v/v 4:3:3) as eluant. Fractions pure by TLC and not positive to Epstein""s spray were combined, concentrated in vacuo to give 1.48 g of the desired product as a white solid. Fractions positive to the Epstein""s spray (contaminated with a small amount of bromide) were combined, concentrated in vacuo, and the residue was triturated with hexanes/ethyl acetate (v/v, 7:3, xcx9c10 ml), and dried in vacuo over P2O5 to afford an additional 0.340 g of the product (total yield: 1.82 g, 73%), mp 165-167xc2x0 C.; 1H-NMR (DMSO-d6) 1.50 (s,9H, But), 2.17 (s, 3H, CH3CO), 3.46 (s, 3H, N3-Me), 4.39 (s, 2H, CH2Cxe2x89xa1C), 4.80 (s, 2H, 6-CH2), 5.22 (s, 2H, 2-CH2), 6.79 (d, J=8.7 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.73 (d, J=8.5 Hz, 2H, 2xe2x80x2,6xe2x80x2-ArH), 7.79, 7.91 (2xc3x97s, 5-H, 8-H); MS (FAB, m/z) 509, 511 [M+, 70%, 30% respectively, Cl isotopic pattern].
Elemental Analysis: Found C, 63.06; H, 5.58; N, 8.13; Cl, 7.08. C27H28ClN3O5 0.25 H2O requires: C, 63.03; H, 5.58; N, 8.17; Cl, 6.81%.
To a solution of tert-butyl 4-[N-[2-acetoxymethyl-7-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.47 g, 0.92 mmol) in tetrahydrofuran (18 ml), was slowly added 1N aqueous NaOH (1.84 ml, 1.84 mmol) followed by water (1.5 ml). The slightly cloudy solution was stirred at room temperature for 1 hour, the solvent was removed in vacuo and the residue was treated with water (35 ml). The pH was adjusted to 4.5 with 1N HCl and the mixture was extracted with ethyl acetate (3xc3x9760 ml). The organics were combined, dried (Na2SO4), and concentrated in vacuo. Purification by column chromatography, on elution with 50% ethyl acetate in dichloromethane, afforded a white solid which was reprecipitated from dichioromethane (minimum amount)/hexanes. The solid was collected by filtration and dried in vacuo over P2O5 (0.345 g, 80%), mp 109-111xc2x0 C.; 1H-NMR (DMSO-d6); 1.49 (s, 9H, CO2But), 3.50 (s, 3H, N3-Me), 3.25 (s (poorly resolved triplet), 1H, Cxe2x89xa1CH), 4.40 (s, 2H, CH2Cxe2x89xa1C), 4.57 (d, J=5.70 Hz, 2H, 2-CH2OH), 4.80 (s, 2H, 6-CH2), 5.67 (t, J=6.4 Hz, 1H, CH2OH), 6.78 (d, J=8.80 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.72 (d, J=8.44 Hz, 2H, 2xe2x80x2,6xe2x80x2-Ar), 7.82, 7.87 (2xc3x97s, 2H, 5-H, 8-H); MS (FAB, m/z) 467, 469 (M+, 95%, 45% respectively; Cl isotopic pattern).
Elemental Analysis: Found: C, 64.44; H, 5.97; N, 8.69; Cl, 7.38. C25H26ClN3O4 requires: C, 64.17; H, 5.60; N, 8.98; Cl, 7.58%.
To a stirred under argon solution of tert-butyl 4-[N-[7-chloro-2-hydroxymethyl-3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.200 g, 0.43 mmol) in anhydrous dichloromethane (5 ml) cooled in an ice-bath was added triethylamine (0.152 g, 1.5 mmol) followed by methanesulphonic anhydride (0.120 g, 0.69 mmol) (added in one portion). After 10 min the ice-bath was removed and stirring was continued for 45 min-TLC (40% ethyl acetate in dichloromethane) indicated a complete reaction. The reaction mixture was then diluted with ethyl acetate (200 ml) and the solution was washed with saturated aqueous sodium bicarbonate (2xc3x9750 ml), and brine (50 ml), dried (Na2SO4) and concentrated in vacuo. Purification by column chromatography, on elution with 40% ethyl acetate in dichloromethane afforded a white solid which dried in vacuo over P2O5 (0.221 g, 94%), mp 204-205xc2x0 C. 1H-NMR (DMSO-d6): 1.50 (s, 9H, CO2But), 3.17 (s (poorly resolved triplet), 1H, Cxe2x89xa1CH), 3.20 (s, 3H, SO2Me), 3.49 (s, 3H, N3-Me), 4.39 (d, J=2.2 Hz, 2H, CH2Cxe2x89xa1C), 4.82 (s, 2H, 6-CH2), 5.41 (s, 2H, 2-CH2), 6.79 (d, J=8.9 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.73 (d, J=8.9 Hz, 2H, 2xe2x80x2,6xe2x80x2-ArH), 7.88, 7.94 (2xc3x97s, 2H, 5-H, 8-H); MS (FAB, m/z) 546, 548 [(M+H)+, 95%, 44% respectively; Cl isotopic pattern].
Elemental Analysis: Found: C, 56.93; H, 5.12; N, 7.50. C26H28ClN3O6S_requires: C, 57.19; H, 5.17; N, 7.70%.
To a solution of tert-butyl 4-[N-[7-chloro-2-methanesulphonyloxymethyl-3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.205 g, 0.38 mmol) in anhydrous dichloromethane (8 ml) under argon was slowly added 1-methylpiperazine (0.376 g, 3.76 mmol). Stirring was continued for 2.5 h at room temperature under argon, then the reaction mixture was diluted with ethyl acetate (200 ml) and washed with 6% Na2CO3 (w/v solution, 2xc3x97100 ml), and dilute brine (100 ml), dried (Na2SO4) and concentrated in vacuo. Purification by column chromatography, on elution with 5% methanol in dichloromethane, afforded a white solid (0.159 g, 77%), mp 136-138xc2x0 C. 1H-NMR (DMSO-d6) 1.50 (s, 9H, But), 2.14 (s, 3H, N-Me piperazine), 2.29 (br s) and 2.50 (brs obscured) (8H, N(CH2CH2)2), 3.60 (s, 3H, N3-Me),3.62 (s, 2H, 2-CH2), 4.38 (d, J=1.1 Hz, 2H, CH2Cxe2x89xa1C), 4.80 (s, 2H, 6-CH2), 6.79 (d, J=8.9 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.73 (d, J=8.9 Hz, 2xe2x80x2,6xe2x80x2-ArH), 7.91, 7.79 (2xc3x97s, 2H, 5-H, 8-H).
MS (FAB, m/z) 550, 552 [(M+H)+, 100%, 35% respectively, Cl isotopic pattern]. Elemental Analysis: Found: C, 65.18; H, 6.58; N, 12.60; Cl, 6.43. C30H36ClN5O3 requires: C, 65.50; H, 6.60; N, 12.73; Cl, 6.44%.
To a stirred under argon solution of tert-butyl 4-[N-[7-chloro-2-methanesulphonyloxymethyl-3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.230 g, 0.42 mmol) in anhydrous dichloromethane (10 ml) was added diethyl amine (0.306 g, 4.2 mmol). The yellow solution was then stirred at room temperature for 18 hours. The solution was then diluted with ethyl acetate (250 ml) and washed with 5% aqueous sodium carbonate solution (2xc3x97100 ml), brine (100 ml), and concentrated in vacuo. Purification by column chromatography, on gradient elution with methanol in dichloromethane (0 to 2%) afforded a white crispy solid which was triturated diethyl ether/hexanes (v/v, 1:2). The white solid was collected by filtration, washed with hexanes and dried in vacuo over P2O5 (0.202 g, 92%), mp 112-113xc2x0 C.; 1H-NMR (DMSO-d6) 0.95 (t, J=6.9 Hz, 6H, 2xc3x97CH2CH3), 1.49 (s, 9H, But), 2.56 (q obscured, J=6.85 Hz, 4H, 2xc3x97CH2CH3), 3.62 (s, 3H, N3-Me), 3.70 (s, 2H, 2-CH2), 4.40 (s, 2H, CH2Cxe2x89xa1C), 4.79 (s, 2H, 6-CH2), 6.78 (d, J=8.9 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.72 (d, J=8.9 Hz, 2xe2x80x2,6xe2x80x2-ArH), 7.81, 7.86 (2xc3x97s, 2H, 5-H, 8-H); MS (FAB, m/z) 523, 525 [(M+H)+, 100%, 40% respectively; Cl isotopic pattern].
Elemental Analysis; Found: C, 65.75; H, 6.67; N, 10.45; Cl, 6.75. C29H35ClN4O3 0.25 H2O requires: C, 66.02; H, 6.78; N, 10.61; Cl, 6.72%.
To a solution of tert-butyl 4-[N-[7-chloro-2-methanesulphonyloxymethyl-3-methyl-4-oxo-3,4-dlhydrbquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.177 g, 0.32 mmol) in anhydrous dichloromethane (7 ml) under argon was added piperidine (0.272 g, 3.2 mmol). The orange solution was stirred at room temperature for 18 hours, then partitioned between ethyl acetate (200 ml) and 5% aqueous sodium carbonate solution (80 ml). The organic layer was washed with 5% aqueous sodium carbonate (80 ml), and dilute brine (80 ml), dried (Na2SO4) and concentrated in vacuo. Purification by column chromatography on gradient elution with ethyl acetate in dichloromethane (5 to 13%) afforded a white solid which was triturated with 5% dichloromethane in hexanes, collected by filtration, washed with hexanes, and dried in vacuo over P2O5 (0.129 g, 75%), mp 162-163xc2x0 C.; 1H-NMR (DMSO-d6) 1.50 (s, 9H, But), 1.42, (br s obscured) and 2.42 (br s) (10H, piperidine 5xc3x97CH2), 3.57 (s, 2H, 2-CH2), 3.61 (s, 3H, N3-Me), 3.20 (s(poorly resolved triplet), 1H, Cxe2x89xa1CH), 4.38 (d, J=2.0 Hz, 2H, CH2Cxe2x89xa1C), 4.79 (s, 2H, 6-CH2), 6.79 (d, J=9.0 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.73 (d, J=8.9 Hz, 2xe2x80x2,6xe2x80x2-ArH), 7.80, 7.89 (2xc3x97S, 2H, 5-H, 8-H); MS (ESI, m/z) 535, 537 [(M+H)+, 100%, 38% respectively; Cl isotopic pattern].
Elemental Analysis: Found: C, 66.92; H, 6.57; N, 10.40; Cl, 6.78. C30H35ClN4O3 requires: C, 67.34; H, 6.59; N, 10.47; Cl, 6.63%.
A mixture of tert-butyl 4-[N-[7-chloro-2-methanesulphonyloxymethyl-3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl )amino]benzoate (0.193 g, 0.35 mmol), dichloromethane (7 ml), and morpholine (0.304 g, 3.5 mmol) was stirred at room temperature for 6 h under argon. The cloudy reaction mixture was then partitioned between ethyl acetate (200 ml) and 5% aqueous sodium carbonate (80 ml). The organic layer was washed more 5% sodium carbonate (2xc3x9780 ml), and brine (80 ml), dried (Na2SO4) and concentrated in vacuo. Purification by column chromatography, on elution with 10% ethyl acetate in dichloromethane, afforded a solid which was triturated with 5% dichloromethane in hexanes. The white solid was collected by filtration, dried in vacuo over P2O5 (0.174 g, 82%), mp 140-143xc2x0 C.; 1H-NMR (DMSO-d6) 1.50 (s, 9H, But), 2.48, (br s obscured) and 3.55 (br s) (8H, N(CH2CH2)2O), 3.62 (s, 5H, 2-CH2, N3-Me), 3.20 (s(poorly resolved triplet), 1H, Cxe2x89xa1CH), 4.39 (s, 2H, CH2Cxe2x89xa1C), 4.79 (s, 2H, 6-CH2), 6.78 (d, J=8.7 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.72 (d, J=8.7 Hz, 2H, 2xe2x80x2,6xe2x80x2-ArH), 7.80, 7.89 (2xc3x97s, 2H, 5-H, 8-H). MS (ESI, m/z) 537, 539 [(M+H)+, 100%, 38% respectively; Cl isotopic pattern].
Elemental analysis: Found: C, 64.81; H, 6.30; N, 10.27; Cl, 6.79. C29H33N4ClO4 requires: C, 64.86; H, 6.19; N, 10.43; Cl, 6.60%.
To a solution of tert-butyl 4-[N-[7-chloro-2-methanesulphonyloxymethyl-3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.213 g, 0.39 mmol) in anhydrous dichloromethane (8 ml) was added pyrrolidine (0.277 g, 3.9 mmol). The clear solution was stirred at room temperature for 4 hours under argon. The clear solution was then partitioned between ethyl acetate (200 ml) and 5% aqueous sodium carbonate. The organic layer was washed with 5% aqueous sodium carbonate (80 ml), and brine (80 ml), dried (Na2SO4), and concentrated in vacuo. The residue was purified by column chromatography on elution with a gradient of ethyl acetate in dichloromethane (20 to 40%). The crispy solid was dissolved in dichloromethane and hexanes and the solvents were removed in vacuo to leave a white solid (0.149 g, 74%), mp 180-183xc2x0 C. 1H-NMR (DMSO-d6) 1.50 (s, 9H, But), 1.70, (br s, 4H, pyrrolidine CH2CH2), 2.54 (br s, 4H, pyrrolidine 2xc3x97Nxe2x80x94CH2), 3.60 (s, 3H, N3-Me), 3.74 (s, 2H, 2-CH2), 4.38 (s, 2H, CH2Cxe2x89xa1C), 4.79 (s, 2H, 6-CH2), 6.79 (d, J=8.7 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.73 (d, J=8.1 Hz, 2H, 2xe2x80x2,6xe2x80x2-ArH), 7.79, 7.89 (2xc3x97s, 2H, 5-H, 8-H).
MS (FAB, m/z) 521, 523 [(M+H)+, 55%, 20% respectively; Cl isotopic pattern]. Elemental Analysis: Found C, 66.60; H, 6.40; N, 10.68; Cl, 7.00. C29H33ClN4O3 requires: C, 66.85; H, 6.38; N, 10.75; Cl, 6.80.
To a solution of tert-butyl 4-[N-[7-chloro-2-methanesulphonyloxymethyl-3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.120 g, 0.22 mmol) in anhydrous dichloromethane (5 ml) under argon was slowly added 1-ethylpiperazine (0.246 g, 2.2 mmol). Stirring was continued for 3 h at room temperature under argon, then the reaction mixture was diluted with ethyl acetate (200 ml) and washed with 5% Na2CO3 (w/v solution, 2xc3x97100 ml), and dilute brine (100 ml), dried (Na2SO4) and concentrated in vacuo. Purification by column chromatography, on elution with 5% methanol in dichloromethane, afforded a white solid (0.086 g, 70%), mp 145-146xc2x0 C. 1H-NMR (DMSO-d6) 0.96 (t, J=6.8 Hz, 3H, CH2CH3), 1.49 (s, 9H, But), 2.29 (q obscured, J=7.2 Hz, 2H, CH2CH3), 2.32 (br s) and 2.48 (brs obscured) (8H, N(CH2CH2)2), 3.29 (s, 1H, Cxe2x89xa1CH), 3.60 (s, 5H, N3-Me, 2-CH2), 4.39 (s, 2H, CH2Cxe2x89xa1C), 4.80 (s, 2H, 6-CH2), 6.78 (d, J=8.3 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.72 (d, J=8.1 Hz, 2xe2x80x2,6xe2x80x2-ArH), 7.81, 7.87 (2xc3x97s, 2H, 5-H, 8-H).
MS (ESI, m/z) 564, 566 [(M+H)+, 100%, 40% respectively, Cl isotopic pattern]. Elemental Analysis: Found: C, 65.05; H, 6.73; N, 12.12; Cl, 6.12. C31H38ClN5O3 0.5H2O requires: C, 64.97; H, 6.86; N, 12.22; Cl, 6.19%.
To a solution of tert-butyl 4-[N-[7-chloro-2-methanesulphonyloxymethyl-3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.150 g, 0.27 mmol) in anhydrous dichloromethaiic (10 ml) under argon was added L-proline methyl ester hydrochloride(0.447 g, 2.70 mmol) followed by triethylamine (0.420 ml, 2.70 mmol). Stirring was continued for 16 h at room temperature under argon, then the reaction mixture was diluted with ethyl acetate (300 ml) and washed with saturated aqueous NaHCO3 solution (2xc3x97100 ml). The aqueous phase was back extracted with ethylacetate (2xc3x9730 ml) and the combined organic phase was washed with brine (50 ml), dried (MgSO4) and concentrated in vacuo. Purification by gradient column chromatography, on elution with 0% to 3% methanol in chloroform, afforded a light yellow solid, which was washed with hexanes (2xc3x9720 ml), the solid removed by filtration and dried in vacuo to afford a white solid (0.132 g, 84%), mp 69-70xc2x0 C. 1H-NMR (DMSO-d6) 1.49 (s, 9H, But), 1.76 (m, 3H, proline Nxe2x80x94CH2xe2x80x94CH2CH), 2.12 (m, 1H, proline Nxe2x80x94CH2CH2xe2x80x94CH), 2.49 (m, 1H, (obscured by DMSO), proline-CH, 2.98 (m, 1H, proline-CH), 3.25 (s, 1H, HCxe2x89xa1C), 3.31 (m, 1H, (obscured by H2O), proline Nxe2x80x94CHCOOCH3), 3.37 (s, 3H, COOCH3), 3.61 (s, 3H, N3-Me), 3.85 (AB system, J=13.2 Hz, 2H, 2-CH2), 4.40 (s, 2H, CH2Cxe2x89xa1C), 4.79 (s, 2H, 6-CH2), 6.77 (d, J=8.9 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.71 (d, J=8.9 Hz, 2H, 2xe2x80x2,6xe2x80x2-ArH), 7.78, 7.87 (2xc3x97s, 2H, 5-H, 8-H).
MS (FAB, m/z) 579, 561, [(M+H)+, 50%, 20% respectively, Cl isotopic pattern]. FAB-HRMS; measured 579.2360; calculated for C31H36ClN4O5 (M+H)+: 579.2374.
To a solution of tert-butyl 4-[N-[7-chloro-2-methanesulphonyloxymethyl-3-methyl-4-oxo-3,4-dihydrquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.220 g, 0.4 mmol) in anhydrous dichloromethane (4 ml) under argon was added a solution of 1-(2-hydroxyethyl)piperazine (0.520 g, 4.0 mmol) in anhydrous dichloromethane (4 ml). Stirring was continued for 5.5 h at room temperature under argon, then the reaction mixture was diluted with ethyl acetate (200 ml) and washed with 5% aqueous sodium carbonate (w/v solution, 2xc3x97100 ml), and dilute brine (100 ml), dried (Na2SO4) and concentrated in vacuo. Purification by column chromatography, on elution with a gradient of methanol in dichloromethane (5 to 10%), afforded a white solid (0.210 g, 90%), mp greater than 90xc2x0 C. (softens). 1H-NMR (DMSO-d6) 1.49 (s, 9H, But), 2.35 (t(obscured), J=6.4 Hz, 2H, Nxe2x80x94CH2CH2OH), 2.37, 2.45 (obscured by DMSO peak), (2xc3x97br s, (8H, N(CH2CH2)2), 3.21 (s, 1H, Cxe2x89xa1CH), 3.46 (q, J=6.11 Hz, 2H, Nxe2x80x94CH2CH2OH), 3.60 (s, 5H, N3-Me, 2-CH2), 4.29 (t, J=5.34 Hz, 1H, Nxe2x80x94CH2CH2OH), 4.39 (s, 2H, CH2Cxe2x89xa1C), 4.80 (s, 2H, 6-CH2), 6.78 (d, J=8.3 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.73 (d, J=8.9 Hz, 2xe2x80x2,6xe2x80x2-ArH), 7.81, 7.89 (2xc3x97s, 2H, 5-H, 8-H).
MS (ESI, m/z) 580, 582 [(M+H)+, 100%, 35% respectively, Cl isotopic pattern].
To a solution of tert-butyl 4-[N-[7-chloro-2-methanesulphonyloxymethyl-3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.197 g, 0.36 mmol) in anhydrous dichloromethane (8 ml) under argon was added 1-phenylpiperazine (0.583 g, 3.6 mmol). Stirring was continued for 2.5 h at room temperature under argon, then the reaction mixture was diluted with ethyl acetate (200 ml) and washed with 5% aqueous sodium carbonate (w/v solution, 2xc3x97100 ml), and dilute brine (100 ml), dried (Na2SO4) and concentrated in vacuo. Purification by column chromatography, on elution with 10% ethyl acetate in dichloromethane, afforded a white solid (0.181 g, 82%), mp greater than 95xc2x0 C. 1H-NMR (DMSO-d6) 1.50 (s, 9H, But), 2.63 (br s) and 3.12 (br s) (8H, N(CH2CH2)2), 3.21 (s, 1H, Cxe2x89xa1CH), 3.64 (s, 3H, N3-Me), 3.70 (s, 2H, 2-CH2), 4.40 (s, 2H, CH2Cxe2x89xa1C), 4.81 (s, 2H, 6-CH2), 6.79 (d, J=8.9 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 6.90 (d, J=8.5 Hz, 2H), 7.20 (t, J=7.8 Hz, 2H), 6.77 (t (obscured), C6H5xe2x80x94N(CH2CH2)2Nxe2x80x94), 7.73 (d, J=8.8 Hz, 2H, 2xe2x80x2,6xe2x80x2-ArH), 7.82, 7.90 (2xc3x97s, 2H, 5-H, 8-H).
MS (ESI, m/z) 612, 614 [(M+H)+, 100%, 40% respectively, Cl isotopic pattern]. Elemental Analysis: Found: C, 68.25; H, 6.28; N, 11.26; Cl, 5.89. C35H38ClN5O3 requires: C, 68.67; H, 6.26; N, 11.44; Cl, 5.79%.
To a solution of tert-butyl 4-[N-[7-chloro-2-methanesulphonyloxymethyl-3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.230 g, 0.42 mmol) in anhydrous dichloromethane (4 ml) under argon was added a solution of 4-hydroxypiperidine (0.424 g, 4.2 mmol) in anhydrous CH2Cl2 (4 ml). The clear solution was stirred at room temperature for 1.5 hours under argon, then partitioned between ethyl acetate (200 ml) and 5% aqueous sodium carbonate solution (100 ml). The organic layer was washed with 5% aqueous sodium carbonate (100 ml), and dilute brine (100 ml), dried (Na2SO4) and concentrated in vacuo. The residue was triturated with ether to give a white precipitate, which was collected by filtration , washed with ether and dried in vacuo over P2O5 (0.206 g, 89%), mp 208-210xc2x0 C.; 1H-NMR (DMSO-d6) 1.50 (s, 9H, But), 1.33 (m, 2H), 1.65 (m, 2H), 2.17 (m, 2H), 2.72 (m, 2H), and 3.50 (m, 1H), (piperidine ring protons), 3.24 (s, 1H, Cxe2x89xa1CH), 3.57, 3.60 (2xc3x97s, 5H, N3-Me , 2-CH2), 4.39 (s, 2H, CH2Cxe2x89xa1C), 4.55 (d, J=4.1 Hz, 1H, OH), 4.79 (s, 2H, 6-CH2), 6.78 (d, J=8.9 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.72 (d, J=8.9 Hz, 2xe2x80x2,6xe2x80x2-ArH), 7.81, 7.87 (2xc3x97s, 2H, 5-H, 8-H); MS (ESI, m/z) 551, 553 [(M+H)+, 100%, 37% rcspectively; Cl isotopic pattern].
Elemental Analysis: Found: C, 62.39; H, 6.27; N, 9.41. C30H35ClN4O4 1.5H2O requires: C, 62.33; H, 6.62; N, 9.69%.
To a suspension of 2-acetoxymethyl-7-chloro-6-methyl-3,4-dihydroquinazolin-4-one (2.00 g, 7.5 mmol) in anhydrous carbon tetrachloride (120 ml) was added N-bromosuccinimide (1.47 g, 8.3 mmol) followed by dibenzoyl peroxide (7.0 mg) under argon. The reaction mixture was placed in a preheated oil bath at 120xc2x0 C. and stirred at this temperature for 3.5 hours while illuminating. The solvent was removed in vacuo and the residue was twice purified by column chromatography using 40% ethyl acetate in chloroform as eluant (1.02 g, 40%), mp 190-195xc2x0 C., 1H-NMR (DMSO-d6) 2.14 (s, 3H, CH3CO), 4.91, 4.97 (2xc3x97s, 4H, 2-CH2 and 6-CH2), 7.78 (s, 1H, 8-H), 8.36 (s, 1H, 5-H), 12.61 (s, 1H, N3xe2x80x94H). This product was used in the next experiment without any further purification.
To a stirred solution of 2-acetoxymethyl-6-bromomethyl-7-chloro-3,4-dihydroquinazolin-4-one (1.02 g, 3.0 mmol) in anhydrous DMF (100 ml) was added tert-butyl 4-N-(prop-2-ynyl)aminobenzoate (0.78 g, 3.4 mmol) followed by 2,6-lutidine (1.23 ml, 10.6 mmol). The reaction mixture was placed in a preheated oil bath at 120xc2x0 C. and stirred at this temperature for 16 hours under argon, then it was allowed to cool to room temperature. The solvent was removed in vacuo and the residue was partitioned between ethyl acetate (300 ml) and half saturated brine (300 ml). The aqueous layer was extracted with more ethyl acetate (2xc3x97100 ml); the combined organic extracts were washed with brine (100 ml), dried (MgSO4) and concentrated in vacuo. Purification by column chromatography on elution with 30% ethyl acetate in chloroform afforded a white solid (0.713 g, 48%), mp 219-220xc2x0 C.; 1H-NMR (DMSO-d6) 1.49 (s, 9H, But), 2.12 (s, 3H, CH3CO), 3.25 (s, 1H, Cxe2x89xa1CH), 4.40 (s, 2H, CH2Cxe2x89xa1C), 4.78 (s, 2H, 6-CH2), 4.94 (s, 2H, 2-CH2), 6.78 (d, J=8.8 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.72 (d, J=8.8 Hz, 2H, 2xe2x80x2,6xe2x80x2-ArH), 7.81, 7.83 (2xc3x97s, 5-H, 8-H), 12.52 (s, 1H, N3xe2x80x94H); FAB-HRMS: measured: 495.1551; calculated for C26H26N3ClO5 495.1561.
Elemental Analysis: Found: C, 62.98; H, 5.27; N, 8.43. C26H26ClN3O5 requires: C, 62.97; H, 5.28; N, 8.47%.
To a solution of tert-butyl 4-[N-[2-acetoxymethyl-7-chloro-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.070 g, 0.14 mmol) in THF (2.7 ml) was dropwise added aqueous NaOH (1N, 0.27 ml, 0.27 mmol) followed by water (0.2 ml). The reaction mixture was stirred at room temperature for 2 hours, then the THF was removed in vacuo. The residue was suspended in water (10 ml) and the pH was adjusted to xcx9c5 with 1N HCl. The white precipitate was. collected by filtration, dried in vacuo and then it was reprecipitated from dichloromethane/hexanes to afford the title compound as a white solid (0.044 g, 70%), mp 185-187xc2x0 C.; 1H-NMR (DMSO-d6) 1.49 (s, 9H, But), 3.25 (s, 1H, Cxe2x89xa1CH), 4.35, 4.38 (2xc3x97s, 4H, CH2Cxe2x89xa1C and 2-CH2), 4.78 (s, 2H, 6-CH2), 5.62 (br s, 1H, OH), 6.78 (d, J=8.7 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.73 (d, J=8.6 Hz, 2H, 2xe2x80x2,6xe2x80x2-ArH), 7.78, 7.84 (2xc3x97s, 2H, 5-H, 8-H), 12.07 (s, 1H, N3xe2x80x94H); FAB-HRMS: measured: 453.1463, calculated for C24H24ClN3O4: 453.1455.
Elemental Analysis: Found: C, 62.50; H, 5.33; N, 9.26. C24H24ClN3O4 0.25H2O requires: C, 62.88; H, 5.38; N, 9.16%.
To a solution of tert-butyl 4-[N-[7-chloro-2-hydroxymethyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.250 g, 0.55 mmol) in anhydrous DMF (6 ml) under argon was added methanesulphonic anhydride (0.191 g, 1.10 mmol) followed immediately by triethylamine (0.27 ml, 1.93 mmol). The clear solution was stirred at room temperature for 45 min then it was partitioned between ethyl acetate (200 ml) and saturated aqueous sodium bicarbonate (60 ml). The organic layer was washed with more saturated aqueous sodium bicarbonate (60 ml), brine (60 ml), dried (Na2SO4) and concentrated in vacuo. Purification by column chromatography on elution with 40% ethyl acetate in dichloromethane afforded a white solid (0.212 g, 73%), mp 178-181xc2x0 C. 1H-NMR (DMSO-d6): 1.49 (s, 9H, CO2But), 3.23 (s, 1H, Cxe2x89xa1CH), 3.20 (s, 3H, SO2Me), 4.39 (s, 2H, CH2Cxe2x89xa1C), 4.78 (s, 2H, 6-CH2), 5.11 (s, 2H, 2-CH2), 6.78 (d, J=9.0 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.72 (d, J=8.8 Hz, 2H, 2xe2x80x2,6xe2x80x2-ArH), 7.85, 7.88 (2xc3x97s, 2H, 5-H, 8-H); MS (ESI, m/z) 554, 556 [(M+Na)+, 100%, 38% respectively; Cl isotopic pattern].
Elemental Analysis: Found: C, 56.66; H, 4.91; N, 7.90. C25H26ClN3O6S_requires: C, 56.44; H, 4.93; N, 7.90%.
To a solution of tert-butyl 4-[N-[7-chloro-2-methanesulphonyloxymethyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.201 g, 0.38 mmol) in anhydrous DMF (5 ml) was added piperidine (0.323 g, 3.8 mmol) and the clear solution was stirred at room temperature for 2.5 hours. The reaction mixture was then partitioned between ethyl acetate (200 ml) and 5% aqueous sodium carbonate (70 ml). The organic layer was washed with 5% aqueous sodium carbonate (70 ml), brine (100 ml), dried (Na2SO4), and concentrated in vacuo. Purification by column chromatography on elution with ethyl acetateldichloromethane (v/v, 1:1) afforded a white solid (0.179 g, 91%), mp 208-210xc2x0 C.; 1H-NMR (DMSO-d6) 1.36 (m), 1.48 (m(obscured)), 1.49 (s) (15H, tBu and piperidine CH2H2CH2) 2.42, (br s, 4H, piperidine CH2NCH2), 3.61 (s, 2H, 2-CH2), 3.22 (s, 1H, Cxe2x89xa1CH), 4.38 (s, 2H, CH2Cxe2x89xa1C), 4.77 (s, 2H, 6-CH2), 6.77 (d, J=9.0 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.72 (d, J=8.9 Hz, 2xe2x80x2,6xe2x80x2-ArH), 7.79, 7.83 (2xc3x97s, 2H, 5-H, 8-H), 11.96 (s, 1H, N3xe2x80x94H).; MS (ESI, m/z) 521, 523 [(M+H)+, 100%, 35% respectively; Cl isotopic pattern].
Elemental Analysis: Found: C, 66.71; H, 6.41; N, 10.50; Cl, 6.85. C29H33ClN4O3 requires: C, 66.85; H, 6.38; N, 10.75; Cl, 6.80%.
To a solution of tert-butyl 4-[N-[7-chloro-4-oxo-2-(piperidin-1-yl)methyl-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.088 g, 0.17 mmol) (Preparation Example 28) in anhydrous DMF (2 ml) was added a sodium hydride (60% dispersion in mineral oil, 8.2 mg, 0.2 mmol).in one portion. The reaction mixture was stirred at room temperature for 3 min under argon; then a solution of N,N-diethylbromoacetamide (N. L. Drake et al, J. Amer. Chem. Soc. 1948, 70, 677-680; 0.066 g, 0.36 mmol) in anhydrous DMF (0.4 ml) was added. The clear solution was stirred at room temperature for 2.5 hours then partitioned between ethyl acetate (40 ml) and brine (40 ml). The aqueous layer was extracted with more ethyl acetate (2xc3x9730 ml) and the combined ethyl acetate extracts were washed with brine (30 ml), dried (Na2SO4) and concentrated in vacuo. Purification by column chromatography on elution with a gradient of ethyl acetate in hexane (20 to 50%) afforded a white solid (0.081 g, 76%), mp 95-97xc2x0 C.; 1H-NMR (CDCl3) 1.13, 1.33 (2xc3x97t, J=7.1 Hz, 6H, 2xc3x97CH2CH3), 1.42 (m obscured, 6H, piperidine CH2CH2CH2), 1.55 (s, 9H, tBu), 2.27 (s, 1H, Cxe2x89xa1CH), 2.42 (m, 4H, piperidine CH2NCH2), 3.41 (m, 4H, 2xc3x97CH2CH3), 3.52 (s, 2H, 2-CH2), 4.17 (s, 2H, CH2Cxe2x89xa1C), 4.75 (s, 2H, 6-CH2), 5.30 (s, 2H, N3xe2x80x94CH2), 6.73 (d, J=9.0 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.78, 8.08 (2xc3x97s, 2H, 5-H, 8-H), 7.86 (d, J=9.02 Hz, 2H, 2xe2x80x2,6xe2x80x2-ArH);); MS (ESI, m/z) 634, 636 [(M+H)+, 100%, 37% respectively; Cl isotopic pattern].
Elemental Analysis: Found: C, 65.88; H, 7.16; N, 10.53. C35H44ClN5O4 requires: C, 66.28; H, 6.99; N. 11.04%.
To a solution of tert-butyl 4-[N-[7-chloro-4-oxo-2-(piperidin-1-yl)methyl-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.052 g, 0.10 mmol) in anhydrous DMF (1 ml) was added sodium hydride (60% dispersion in mineral oil, 5.00 mg, 0.12 mmol) in one portion. The reaction mixture was stirred at room temperature for 3 min under argon; then a solution of 1-(bromoacetyl)piperidine (M. Arimoto et al. The Journal of Antibiotics 1986, 1243-1256; 0.041 g, 0.2 mmol) in anhydrous DMF (0.2 ml) was added. The clear solution was stirred at room temperature for 3 hours then partitioned between ethyl acetate (40 ml) and brine (40 ml). The aqueous layer was extracted with ethyl acetate (2xc3x9720 ml) and the combined ethyl acetate extracts were washed with brine (30 ml) dried (Na2SO4) and concentrated in vacuo. Purification by column chromatography on elution with a gradient of ethyl acetate in hexanes (40 to 50%) afforded a white solid (0.040 g, 62%), mp greater than 104xc2x0 C.; 1H-NMR (DMSO-d6) 1.30-1.65 (m, 12H), 2.36 (br s, 4H) and 3.40 (m, 4H), (piperidine protons), 1.49 (s, 9H, tBu), 3.21 (s, 1H, Cxe2x89xa1CH), 3.47 (s, 2H, 2-CH2), 4.39 (s, 2H, CH2Cxe2x89xa1C), 4.80 (s, 2H, 6-CH2), 5.10 (s, 2H, N3xe2x80x94CH2), 6.78 (d, J=8.9 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.83, 7.84 (2xc3x97s, 2H, 5-H, 8-H), 7.73 (d, J=8.8 Hz, 2H, 2xe2x80x2,6xe2x80x2-ArH); MS (ESI, m/z) 646, 648 [(M+H)+, 100%, 36% respectively; Cl isotopic pattern].
Elemental Analysis: Found: C, 65.84; H, 6.69; N, 10.42. C36H44ClN5O4 0.5H2O requires: C, 65.99; H, 6.92; N, 10.68%.
To a solution of of tert-butyl 4-[N-[7-chloro-4-oxo-2-piperidin-1-ylmethyl-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.096 g, 0.18 mmol) in anhydrous DMF (5 ml) under argon was added sodium hydride (60% dispersion in mineral oil, 8 mg, 0.2 mmol) in one portion. The reaction mixture was stirred at room temperature for 3 min, then methyl bromoacetate (0.141 g, 0.9 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 24 hours then partitioned between ethyl acetate (150 ml) and half saturated brine (100 ml). The organic layer was washed with more brine (100 ml). The combined aqueous washings were extracted with ethyl acetate (2xc3x9750 ml). The combined ethyl acetate extracts were washed with brine (100 ml), dried (Na2SO4) and concentrated in vacuo. Purification by column chromatography on elution with chloroform afforded a white solid (0.071 g, 67%), mp 148-150xc2x0 C.; 1H-NMR (DMSO-d6) 1.36 (m, 6H, piperidine CH2CH2CH2), 1.50 (s, 9H, But), 2.32, (m, 4H, piperidine CH2NCH2), 3.23 (s, 1H, Cxe2x89xa1CH), 3.58 (s, 2H, 2-CH2), 3.67 (s, 3H, CO2Me), 4.40 (d, J=2.0 Hz, 2H, CH2Cxe2x89xa1C), 4.80 (s, 2H, 6-CH2), 4.87 (s, 2H, N3xe2x80x94CH2), 6.77 (d, J=9.0 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.72 (d, J=8.9 Hz, 2xe2x80x2,6xe2x80x2-ArH), 7.86, 7.87 (2xc3x97s, 2H, 5-H, 8-H); MS (FAB, m/z) 593, 595 [(M+H)+, 100%, 36% respectively; Cl isotopicpattern].
FAB-HRMS: measured: 593.2506; calculated for C32H38ClN4O5 593.2531.
To a solution of of tert-butyl 4-[N-[7-chloro-4-oxo-2-piperidin-1-ylmethyl-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]benzoate (0.250 g, 1.69 mmol) in anhydrous DMF (10 ml) under argon was added lithium chloride (0.071 g, 6.24 mmol). When the lithiumn chloride had dissolved the reaction mixture was cooled to 4xc2x0 C. in an ice-bath and then sodium hydride (60% dispersion in mineral oil, 21 mg, 0.52 mmol) in one portion followed by 2-dimethylaminoethyl chloride (free base, 0.690 g, 6.40 mmol). Stirring was continued at 4xc2x0 C. for 15 min and then for an additional 16 h at room temperature. The reaction mixture was then heated to 70xc2x0 C. and then more sodium hydride (60% dispersion in mineral oil, 10 mg, 0.25 mmol) was added followed by 2-dimethylaminoethyl chloride (free base, 0.690 g, 6.40 mmol). Stirring was continued at this temperature for 2 hours under argon; the the solvent was removed in vacuo and the residue was partitioned between ethyl acetate (150 ml) and water (200 ml). The organic layer was washed with more water (150 ml). The aqueous washings were extracted with more ethyl acetate (2xc3x9750 ml). The combined ethyl acetate extracts were washed with brine ((50 ml), dried (Na2SO4) and concentrated in vacuo. Purification by column chromatography on elution with ether/hexanes/MeOH (v/v/v: 5:4: 1) afforded a white solid (0.074 g, 26%), mp 75-77xc2x0 C.; 1H-NMR (DMSO-d6) 1.35-1.49 (m, 15H, tBu, piperidine CH2CH2CH2), 2.19 (s, 6H, NMe2), 2.40 (m, 4H, piperidine CH2NCH2), 2.53 (t (obscured), 2H, Me2NCH2), 3.27 (s, 1H, Cxe2x89xa1CH), 3.61 (s, 2H, 2-CH2), 4.23 (t, J=7.0 Hz, 2H, N3xe2x80x94CH2), 4.40 (s, 2H, CH2Cxe2x89xa1C), 4.79 (s, 2H, 6-CH2), 6.77 (d, J=9.0 Hz, 2H, 3xe2x80x2,5xe2x80x2-ArH), 7.72 (d, J=8.9 Hz, 2xe2x80x2,6xe2x80x2-ArH), 7.82, 7.87 (2xc3x97s, 2H, 5-H, 8-H); MS (ESI, m/z) 592, 594 [(M+H)+, 100%, 38% respectively; Cl isotopic pattern].
Elemental Analysis: Found: C, 66.45; H, 7.12; N, 11.41; C33H42ClN5O3 0.25H2O requires: C, 66.42; H, 7.13; N, 11.73%.
Anhydrous methanol (36 ml) was added into a 250 ml round-bottomed flask that contained sodium (0.081 g) under argon. To this stirred solution chloroacetonitrile (1.56 g, 20.7 mmol) was added and stirring was continued for 30 min at room temperature under argon. 5-Amino-6-carboxyindane (European Patent Application 0602851A1; 3.19 g, 18.0 mmol) was then added and the reaction mixture was diluted with more methanol (42 ml). After the reaction mixture being stirred for 1 hour at room temperature, more anhydrous methanol (25 ml) was added and stirring was continued for 1 hour at room temperature and then for a further 1 hour at 80xc2x0 C. under argon. The reaction mixture was then allowed to cool to room temperature; the white precipitate was collected by filtration, washed with methanol, water and dried in vacuo over P2O5 (3.15 g, 75%), mp 270-272xc2x0 C.; 1H-NMR (DMSO-d6) 2.07 (m, 2H, 7-CH2), 2.98 (m, 4H, 6-CH2 and 8-CH2), 4.52 (s, 2H, 2-CH2), 7.50 (s, 1H, 9-H), 7.93 (s, 1H, 5-H), 12.37, (s, 1H, N3xe2x80x94H). MS (FAB, m/z) 235 (M+H)+;
Elemental Analysis: Found: C, 61.46; H, 4.84; N, 12.03; Cl, 14.91%. C12H11N2ClO requires: C, 61.42; H, 4.72; N, 11.94; Cl, 15.11%.
A solution of powdered and dried cesium acetate (41.00 g, 213.5 mmol) in DMF (150 ml) was heated at 60xc2x0 C. and stirred at this temperature for 30 min under argon. The reaction mixture was cooled to 40xc2x0 C. and then a solution of 2-chloromethyl-3,4,7,8-tetrahydro-6H-cyclopenta[g]quinazolin-4-one (10.00 g, 42.7 mmol) in DMF (350 ml) was added via a cannula under argon. The reaction mixture was heated to 80xc2x0 C. and stirred for 3 hours at this temperature. The solvent was then removed in vacuo and the residue was stirred with chloroform (500 ml), then it was filtered through a celite. The celite was washed with chloroform (2xc3x97100 ml); the combined organics were washed with water (2xc3x97150 ml), brine (100 ml), dried (MgSO4) and concentrated in vacuo to leave a beige solid. Recrystallisation from toluene (twice) afforded a white solid (4.4 g, 40%) mp greater than 200xc2x0 C. (dec); 1H-NMR (DMSO-d6) 2.04 (m, 2H, 7-CH2), 2.12 (s, 3H, CH3CO), 2.98 (m, 4H, 6-CH2 and 8-CH2), 4.94 (s, 2H, 2-CH2), 7.47 (s, 1H, 9-H), 7.92 (s, 1H, 5-H), 12.26, (s, 1H, N3xe2x80x94H). MS (ESI, m/z) 217 (M-CH3CO)+.
To a solution of 2-acetoxymethyl-3,4,7,8-tetrahydro-6H-cyclopenta[g]quinazolin-4-one (1.00 g, 3.9 mmol) in DMA (45 ml) was added powdered potassium carbonate (1.35 g, 9.8 mmol). The reaction mixture was stirred for 1 hour at room temperature under argon, then it was placed in an ice-bath and chloromethyl pivalate (0.880 g, 5.8 mmol) was added dropwise. The ice-bath was removed and stirring was continued for a further 16 hours under argon. The solvent was removed in vacuo and the residue was partitioned between ethyl acetate (50 ml) and water (50 ml). The aqueous layer was extracted with more ethyl acetate (3xc3x9730 ml). The ethyl acetate extracts were combined, washed with brine (40 ml), dried (MgSO4) and concentrated in vacuo. Purification by column chromatography using 40% ether in hexane as eluant afforded in order of elution: 2-acetoxymethyl-4-pivaloyloxymethyloxy-7,8-tetrahydro-6H-cyclopenta[g]quinazoline as a colourless oil (0.875 g, 60%); 1H-NMR (DMSO-d6) 1.10 (s, 9H, POM C(CH)3)3), 2.08 (m, 2H, 7-CH2), 2.16 (s, 3H, CH3CO), 2.98 (m, 4H, 6-CH2 and 8-CH2), 5.20 (s, 2H, 2-CH2), 6.23 (s, 2H, POM CH2), 7.72, 7.88 (2xc3x97s, 2H, 9-H, 5-H); MS (ESI, m/z) 373 (M+H)+;
Elemental Analysis: Found: C, 64.45; H, 6.54; N, 7.41. C20N24N2O5 requires: C, 64.50; H, 6.50; N, 7.52%.
2-acetoxymethyl-3-pivaloyloxymethyl-3,4,7,8-tetrahydro-6H-cyclopenta[g]quinazolin-4-one as a solid (0.380 g, 26%), mp 132-135xc2x0 C.; 1H-NMR (DMSO-d6) 1.12 (s, 9H, POM C(CHd3)3), 2.07 (m, 2H, 7-CH2), 2.16 (s, 3H, CH3CO), 2.98 (m, 4H, 6-CH2 and 8-C2), 5.25 (s, 2H, 2-CH2), 6.03 (s, 2H, POM CH2), 7.50 (s, 1H, 9-H) 7.96 (s, 1H, 5-H); MS (ESI, m/z) 373 (M+H)+;
Elemental Analysis: Found: C, 64.43; H, 6.53; N, 7.54. C20N24N2O5 requires: C, 64.50; H, 6.50; N, 7.52%.
To a vigorously stirred suspension of chromium(VI) oxide (0.040 g) in dichloromethane (15 ml) cooled in an ice-bath was added tert-butyl hydroperoxide (70% aqueous solution, 7.21 g, 56.0 mmol) dropwise. The ice-bath was then removed, the reaction mixture was allowed to stir for 10 min at room temperature and then a solution of 2-acetoxymethyl-3-pivaloyloxymethyl-3,4,7,8-tetrahydro-6H-yclopenta[g]quinazolin-4-one (1.50 g, 4.0 mmol) in dichloromethane (10 ml) was added dropwise. The red reaction mixture was stirred for 24 hours at room temperature then it was cooled in an ice-bath and then 10% aqueous sodium metabisulfite solution (20 ml) was added dropwise. The reaction mixture was allowed to stir for 2 hours at room temperature; then it was partitioned between ethyl acetate (50 ml) and half-saturated brine (50 ml). The aqueous layer was extracted with more ethyl acetate (3xc3x9730 ml). The ethyl acetate extracts were combined, washed with saturated aqueous sodium bicarbonate (50 ml), brine (50 ml), dried (MgSO4) and concentrated in vacuo. Purification by column chromatography on elution with a gradient of ethyl acetate in hexanes (20 to 50%) afforded in order of elution: 2-acetoxymethyl-3-pivaloyloxymethyl-3,4,7,8-tetralhydro-6H-cyclopenta[g]quinazolin-4,8-dione as a white solid (0.381 g, 25%), mp 144xc2x0 C.; 1H-NMR (CDCl3) 1.13 (s, 9H, POM C(CH3)3), 2.16 (s, 3H, CH3CO), 2.74 (m, 2H, 7-CH2), 3.20 (m, 2H, 6-CH2), 5.16 (s, 2H, 2-CH2), 6.05 (s, 2H, POM CH2), 7.93 (s, 1H, 9-H), 8.30 (s, 1H, 5-H); MS (ESI, m/z) 387 (M+H)+; 2-acetoxymethyl-3-pivaloyloxymethyl-3,4,7,8-tetrahydro-6H-cyclopenta[g]quinazolin-4,6-dione as a white solid (0.697 g, 45%), mp 147xc2x0 C.; 1H-NMR (CDCl3) 1.17 (s, 9H, POM C(CHd3)3), 2.19 (s, 3H, CH3CO), 2.80 (m, 2H, 7-CH2), 3.27 (m, 2H, 8-CH2), 5.19 (s, 2H, 2-CH2), 6.09 (s, 2H, POM CH2), 7.70 (s, 1H, 9-H) 8.66 (s, 1H, 5-H); MS (ESI, m/z) 387 (M+H)+.
2-Acetoxymethyl-3-pivaloyloxymethyl-3,4,7,8-tetrahydro-6H-cyclopenta[g]quinazolin-4,6-dione (0.140 g, 0.36 mmol) was suspended in anhydrous methanol (8 ml) and then anhydrous dichoromethane (4 ml) was added until a clear solution was obtained. Tert-butyl 4-aminobenzoate (0.084 g, 0.44 mmol) was then added followed by decaborane (0.014 g, 0.12 mmol). The reaction mixture was stirred overnight at room temperature under argon. The solvent was removed in vaciio, and the residue was purified by column chromatography on elution with 40% ethyl acetate in hexane. The product was obtained as a glass (0.127 g, 63%) 1H-NMR (CDCl3) 1.20 (s, 9H, POM C(CH3)3), 1.58 (s, 9H, CO2C(CH3)3), 2.20 (s, 3H, CH3CO), 2.02, 2.72 (2xc3x97m, 2H, 7-CH2), 3.13 (m, 2H, 8-CH2), 4.36 (d, J=7.7 Hz, 1H, NH), 5.15 (q, J=6.8 Hz, 1H, 6-H), 5.21 (s, 2H, 2-CH2), 6.12 (AB system, J=10.7 Hz, 2H, POM CH2), 6.67 (d, J=8.8 Hz, 2H, 3,5xe2x80x2-ArH), 7.57 (s, 1H, 9-H), 7.85 (d, J=8.9 Hz, 2H, 2xe2x80x2,6xe2x80x2-ArH), 8.25 (s, 1H, 5-H); MS (ESI, m/z) 564 (M+H)+.
To a stirred solution of tert-butyl 4-[N-[-2-acetoxymethyl-4-oxo-3-pivaloyloxymethyl-3,4,7,8-tetrahydro-6H-cyclopenta[g]quinazolin-6-yl]amino]benzoate (0.060 g, 0.106 mmol) in THF (6 ml) was added glacial acetic acid (2 ml) followed by 37% aqueous formaldehyde (0.090 ml). The solution was stirred for 2.5 hours at room temperature and then sodium cyanoborohydride (0.017 g, 0.27 mmol) was added. The reaction mixture was stirred overnight at room temperature. The solvent was removed in vacuo and the residue was partitioned between ethyl acetate (25 ml) and water (25 ml). The aqueous layer was extracted with more ethyl acetate (2xc3x9725 ml). The combined organics were washed with saturated aqueous sodium bicarbonate (30 ml), brine (30 ml), dried (Na2SO4) and concentrated in vacuo to leave an oil (0.060 g, 98%); 1H-NMR (CDCl3) 1.20 (s, 9H, POM C(CH3)3), 1.58 (s, 9H, CO2C(CH3)3), 2.15 (s, 3H, CH3CO), 2.02, 2.52 (2xc3x97m, 2H, 7-CH2), 2.74 (s, 3H, N10-Me), 3.13 (m, 2H, 8-CH2) 5.21 (s, 2H, 2-CH2), 5.66 (t, 1H, J=8.1 Hz, 6-H), 6.12 (AB system, J=10.8 Hz, 2H, POM CH2), 6.85 (d, J=8.8 Hz, 2H, 3,5xe2x80x2-ArH), 7.58 (s, 1H, 9-H), 7.90 (d, J=8.9 Hz, 2H, 2,6xe2x80x2-ArH), 8.08 (s, 1H, 5-H); MS (ESI, m/z) 578 (M+H)+.
To a stirred solution of tert-butyl 4-[N-[-2-acetoxymethyl-4-oxo-3-pivaloyloxymethyl-3,4,7,8-tetrahydro-6H-cyclopenta[g]quinazolin-6-yl]-N-methylamino]benzoate (0.060 g, 0.106 mmol) in methanol (2.5 ml) was added aqueous NaOH solution (1M, 0.4 ml, 0.4 mmol) followed by water (0.5 ml). The solution was stirred overnight at room temperature, then the methanol was removed in vacuo, and the pH of the aqueous residue was adjusted to xcx9c4 with 1N HCl to give a milky suspension. This was extracted with ethyl acetate (3xc3x9725 ml); the combined extracts were washed with water (20 ml), brine (20 ml), dried (Na2SO4) and concentrated in vacuo to give an off-white solid (0.045 g, 100%) 1H-NMR (CDCl3) 1.58 (s, 9H, CO2C(CH3)3), 2.04, 2.52 (2xc3x97m, 2H, 7-CH2), 2.75 (s, 3H, N10-Me), 3.13 (m, 2H, 8-CH2), 4.71 (s, 2H, 2-CH2), 5.66 (t, 1H, J=8.1 Hz, 6-H), 6.86 (d, J=9.0 Hz, 2H, 3,5xe2x80x2-ArH), 7.56 (s, 1H, 9-H), 7.89 (d, J=8.9 Hz, 2H, 2,6xe2x80x2-ArH), 8.05 (s, 1H, 5-H); MS (ESI, m/z) 444 [(M+Na)+ , 100%], 422 [(M+H)+, 20%]. This compound was also made from 2-acetoxymethyl-4-pivaloyloxymethyloxy-7,8-tetrahydro-6H-cyclopenta[g]quinazoline by first brominating this compound, followed by the displacement of the bromide with tert-butyl 4-aminobenzote, then methylation and finally removal of the POM and acetyl groups under alkaline conditions.
To a solution of tert-butyl 4-[N-[2-hydroxymethyl-4-oxo-3,4,7,8-tetrahydro-6H-cyclopenta[g]quinazolin-6-yl]-N-methylamino]benzoate (0.130 g, 0.31 mmol) in anhydrous DMF (4 ml) under argon was added methanesulphonic anhydride (0.1 12 g, 0.64 mmol) followed immediately by triethylamine (0.16 ml, 1.12 mmol). The solution was stirred for 45 min after which time the reaction mixture was partitioned between a saturated aqueous solution of sodium bicarbonate (40 ml) and ethyl acetate (75 ml). The organic phase was then successively washed with a saturated aqueous solution of sodium bicarbonate (30 ml) and brine (30 ml), then dried (Na2SO4) and concentrated under reduced pressure. Flash column chromatography with dichloromethane-ethyl acetate (3:2 v/v) afforded the title compound (0.70 g, 45%) 1H NMR (DMSO-d6) 1.51 (s, 9H, C(CH3)3), 2.03, 2.45 (2xc3x97m, 2H, 7-CH2), 2.66 (s, 3H, N10-Me), 3.02 (m, 2H, 8-CH2), 3.34 (s, 3H, xe2x80x94SO2CH3), 5.12 (s, 2H, CH2OMs), 5.78 (t, 1H, J=8.0 Hz, 6-H), 6.96 (d, 2H, J=9.0 Hz, 3xe2x80x2,5xe2x80x2-ArH), 7.58 (s, 1H, 9-H), 7.71 (s, 1H, 5-H), 7.75 (d, 2H, J=9.0 Hz, 2xe2x80x2,6xe2x80x2-ArH); MS (ESI, m/z) 522 [(M+Na)+ 100%, 500 (M+H)+ 7%].
To a solution of tert-butyl 4-[N-[2-(methanesulphonyl)methyl-4oxo-3,4,7,8-tetrahydro-6H-cyclopenta[g]quinazolin-6-yl]-N-methylamino]benzoate (0.070 g, 0.14 mmol) in anhydrous DMF (2.5 mL) was added piperidine (0.14 ml, 1.42 mmol) and the solution was stirred for 2.5 hours. The reaction mixture was partitioned between a saturated aqueous solution of sodium bicarbonate (35 ml) and ethyl acetate (75 ml). The organic phase was successively washed with a 10% (w/v) aqueous solution of sodium carbonate (25 ml), brine (25 ml), then dried (Na2SO4) and concentrated under reduced pressure to afford the title compound (0.067 g, 98%). 1H NMR (CDCl3) 1.54 (m), 1.59 (s), 1.63 (m (obscured)) (15H, tBu and piperidine CH2CH2CH2), 2.12 (m, 1H, 7-CH), 2.54 (m, 5H, 7-CH and piperidine CH2NCH2), 2.75 (s, 3H, N10-Me), 3.09 (m, 2H, 8-CH2), 3.51 (s, 2H, 2-CH2), 5.66 (t, 1H, J 8.2 Hz), 6.85 (d, 2H, J=8.9 Hz, 3,xe2x80x25xe2x80x2-ArH), 7.53 (s, 1H, 9-H), 7.89 (d, 2H, J=8.9 Hz, 2xe2x80x2,6xe2x80x2-ArH), 8.06 (s, 1H, 5-H); MS (ESI, m/z) 489 [(M+H)+ 100%].
To a solution of tert-butyl 4-[N-[4-oxo-2-(piperidin-1-yl)methyl-3,4,7,8-tetrahydro-6H-cyclopenta[g]quinazolin-6-yl]-N-methylamino]benzoate (0.060 g, 0.123 mmol) in anhydrous DMF (3 ml) under argon was added a dispersion of NaH in mineral oil (60% w/w) (0.0060 g, 0.147 mmol) and the reaction mixture was stirred for 1 min. Mel (15 xcexcL, 0.246 mmol) was then added via syringe and the reaction mixture was stirred for a further 2 hours. The reaction mixture was then partitioned between a saturated aqueous solution of sodium bicarbonate (40 ml) and ethyl acetate (75 ml) and the organic phase was then successively washed with a saturated aqueous solution of sodium bicarbonate (30 ml), brine (30 ml), then dried (Na2SO4) and concentrated under reduced pressure to afford the title compound (0.056 g, 91%).
1H NMR (CDCl3) 1H 1.46 (m), 1.59 (s) (15H, tBu and piperidine CH2CH2CH2), 2.07 (m, 1H, 7-CH), 2.57 (m, 5H, 7-CH and piperidine CH2NCH2), 2.73 (s, 3H, N10-Me), 3.08 (m, 2H, 8-CH2), 3.58 (s, 2H, 2-CH2), 3.77 (s, 3H, 3-Me) 5.66 (t, 1H, J=8.1 Hz), 6.85 (d, 2H, J=8.9 Hz, 3,xe2x80x25xe2x80x2-ArH), 7.54 (s, 1H, 9-H), 7.90 (d, 2H, J=8.9 Hz, 2xe2x80x2,6xe2x80x2-ArH), 8.05 (s, 1H, 5-H); MS (ESI, m/z) 503 [(M+H)+ 100%].